Osteopontin/OPN: Novus Biologicals

AF808

Osteopontin/OPN Antibody [Unconjugated]

(2 Reviews)

100 μg

AF808

NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and secreted molecule. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox">

Osteopontin/OPN was detected in immersion fixed C2C12 mouse myoblast cell line (left panel, positive stain) and mouse splenocytes (right panel, negative stain) using Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; <a class=

NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and secreted molecule. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." alt="Osteopontin/OPN was detected in immersion fixed C2C12 mouse myoblast cell line (left panel, positive stain) and mouse splenocytes (right panel, negative stain) using Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and secreted molecule. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_lightbox">

Osteopontin/OPN was detected in perfusion fixed frozen sections of mouse thymus using Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; <a class=

CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_thumb" />

441-OP), immobilized onto a microplate, supports the adhesion of the HEK293 human embryonic kidney cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Mouse Osteopontin/OPN (2 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808). The ND₅₀ is typically 1-3 µg/mL." class="big_lightbox">

Recombinant Mouse Osteopontin/OPN (<a class=

441-OP), immobilized onto a microplate, supports the adhesion of the HEK293 human embryonic kidney cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Mouse Osteopontin/OPN (2 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808). The ND₅₀ is typically 1-3 µg/mL." class="big_thumb" />

Notch ligands Jag1 and Dll1 are both required for segregation of hepatocytic fate centrally and biliary fate peripherally.(A) Immunolabeling for OPN and HNF4A of BMEL cells presented with DLL4 on 30 kPa substrates. Control cells were transduced with an shRNA vector coding for a non-mammalian target. shJag1 and shDll1 cells were transduced with shRNA vectors targeting Jag1 and Dll1, respectively. Scale bar is 150 µm. (B) Confocal imaging of immunolabeled SOX9 and HNF4A in control, shJag1, and shDll1 cells presented with DLL4 on 30 kPa substrates. Scale bar is 75 µm. (C) Quantification of OPN+ cell counts of control, shJag1, and shDll1 cells presented with DLL4 on 30 kPa substrates. (D) Quantification of SOX9 and HNF4A intensity of control, shJag1, and shDll1 cells presented with DLL4 on 30 kPa substrates. (C, D) Mean ± 95% CI.10.7554/eLife.38536.024Figure 7—source data 1.Summary table for OPN data in Figure 7C.10.7554/eLife.38536.025Figure 7—source data 2.Summary table for SOX9 and HNF4A data in Figure 7D.Summary table for OPN data in Figure 7C.Summary table for SOX9 and HNF4A data in Figure 7D.Regression analysis of OPN+ cell counts.Data in Figure 7B were separated into peripheral and central subsets for which dimensionless radius was greater than 0.75 (R>0.75) and less than 0.75 (R<0.75). Separate multiple regression models were generated for each data subset for which coefficient estimates (corresponding to mean change in cell counts) and 95% CI were plotted. For each factor, 95% CI that do not intersect with the dashed line indicate regression coefficient estimates for which P<0.05. Control, shJag1, and shDll1 cells were presented with DLL4 on 30 kPa substrates. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30589410), licensed under a CC-BY license. Not internally tested by R&D Systems.

Altered expression of ECM proteins in Cyp1b1-/- LSEC.Western blot analysis of ECM proteins in the conditioned medium (CM) and cell lysates from LSEC was performed. (A) The levels of SPARC, Tenascin-C, TSP2, and Osteopontin were below the level of detection. Cyp1b1 LSEC produced TSP1, periostin, and fibronectin. (B) The quantitative assessment of the data. TSP1 level was significantly decreased in lysates from Cyp1b1-/- LSEC (***P< 0.001; n = 3), while periostin level was increased in lysates from Cyp1b1-/- LSEC. The level of fibronectin secreted into the conditioned medium was decreased in Cyp1b1-/- LSEC. These experiments were repeated with two isolation of LSEC with similar results. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30372497), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>Peripheral biliary differentiation is dependent on both Notch signaling and substrate stiffness.(A) Immunolabeling for OPN of BMEL cells presented with DLL4 on 30 kPa and 4 kPa substrates. Cells were treated with vehicle control (DMSO) or an inhibitor of Notch signaling ( gamma -secretase inhibitor X, GSI, 5 µM). (B) Quantification of OPN+ cell counts on 30 kPa and 4 kPa substrates after treatment with DMSO or GSI. (C) Immunolabeling for SOX9 and HNF4A of BMEL cells on 30 kPa and 4 kPa substrates. (D) Quantification of SOX9 and HNF4A intensity on 30 kPa and 4 kPa substrates. (E) RNA in situ hybridization for Jag1, Dll1, and Notch2 on 30 kPa and 4 kPa substrates. Cells were exogenously presented with IgG or DLL4. (A, C, E) Scale bars indicate 150 µm. (B, D) Mean ± 95% CI.10.7554/eLife.38536.013Figure 2—source data 1.Summary table for OPN data in Figure 2B and Figure 2—figure supplement 1.10.7554/eLife.38536.014Figure 2—source data 2.Summary table for SOX9 and HNF4A data in Figure 2D.Summary table for OPN data in Figure 2B and Figure 2—figure supplement 1.Summary table for SOX9 and HNF4A data in Figure 2D.Quantification of OPN+ cell counts in arrayed patterns.Cells were cultured on 30 kPa and 4 kPa substrates and presented with IgG, DLL1, DLL4, and JAG1. Treatments included vehicle control (DMSO) or an inhibitor of Notch signaling ( gamma -secretase inhibitor X, GSI, 5 µM).Regression analysis of OPN+ and ALB+ cell counts.Data in Figure 2B were separated into peripheral and central subsets for which dimensionless radius was greater than 0.75 (R>0.75) and less than 0.75 (R<0.75). Separate multiple regression models were generated for each data subset for which coefficient estimates (corresponding to mean change in cell counts) and 95% CI were plotted for OPN+ (A) and ALB+ (B) cells. For each factor, 95% CI that do not intersect with the dashed line indicate regression coefficient estimates for which P<0.05. Cells were cultured on 30 kPa and 4 kPa substrates and presented with IgG,

Characterisation of mouse liver progenitor cell lines.(A) Mouse liver progenitor cell (LPC) lines (BMOL1.2, BMOL-TAT) were stained using immunofluorescence to determine expression of LPC marker Osteopontin (OPN) (red). IgG isotype control images obtained using identical imaging conditions. DAPI, blue. (B) LPCs (BMOL1.2) were shown to express full-length primary cilium (Pc) structures by immunofluorescence detection of axoneme ( alpha -acetylated tubulin, green) and basal body ( gamma -tubulin, red) markers. This was also confirmed by scanning electron microscopy (SEM). BMOL-TAT cells were also confirmed to express Pc via immunofluorescence and EM studies (data not shown). (C) Nuclear GLI2 (red) expression in LPC lines (BMOL1.2, BMOL-TAT) via immunofluorescence staining. DAPI, blue. Confocal microscopy, 63x objective. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0171480), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>Osteopontin (OPN) drives enhancement in macrophage (Mφ) M2 polarization and angiogenic capacity. (A) Representative images of protein expression profiles obtained by comprehensive protein array in each Mφ subset. Red arrowheads indicate OPN. (B) The mRNA expression level of Spp1 relative to glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was analyzed by real-time reverse transcription polymerase chain reaction in each Mφ subset and was normalized to Mφ (–), n = 6 [***p < 0.001 vs. untreated, #p < 0.05 vs. interleukin (IL)-10 alone]. (C) The protein expression level of OPN relative to GAPDH was measured by western blotting and was normalized to Mφ (–), n = 10. Lower panels are typical images of each protein (***p < 0.001 vs. untreated, #p < 0.05 vs. IL-10 alone). (D) Representative confocal laser scanning immunofluorescence overlay images of OPN (red) and DAPI (blue) in each Mφ subset. Scale bar represents 20 µm. Images in the right row are magnified regions from white or yellow rectangles in the panels of corresponding groups. Scale bar represents 10 µm. (E) Relative mean fluorescence intensity (MFI) of CD163 was measured by FACS analysis in each Mφ subset. An anti-OPN antibody (Ab) and its isotype-matched control Ab were used at 3 µg/mL, n = 4 (***p < 0.001 vs. untreated, ##p < 0.01, #p < 0.05 vs. IL-10 alone, †††p < 0.001 vs. IL-10 + IL-18). (F) The total areas and lengths of tube-like structures were determined by the Matrigel tube formation assay where b.End5 was cocultured with each Mφ subset, n = 12 (***p < 0.001, **p < 0.01, *p < 0.05 vs. untreated, #p < 0.05 vs. IL-10 alone, †††p < 0.001 vs. IL-10 + IL-18). All data are expressed as means ± SEM and were analyzed by a one-way ANOVA followed by Tukey’s test. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29559970), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>Thrombin contributes to macrophage (Mφ) M2 polarization and angiogenic capacity through proteolytic modification for osteopontin (OPN). (A) The mRNA expression level of Prothrombin relative to glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was analyzed by reverse transcription polymerase chain reaction in each Mφ subset and were normalized to Mφ (–), n = 7 [***p < 0.001, **p < 0.01 vs. untreated, ##p < 0.01 vs. interleukin (IL)-10 alone]. (B,C) The protein expression levels of (B) thrombin or (C) OPN N-Half relative to GAPDH were measured by western blotting in each Mφ subset and were normalized to Mφ (–). Lower panels are typical images of each protein. (B)n = 8 (***p < 0.001, *p < 0.05 vs. untreated, #p < 0.05 vs. IL-10 alone). (C)n = 16 (***p < 0.001, *p < 0.05 vs. untreated, ##p < 0.01, #p < 0.05 vs. IL-10 alone, †††p < 0.001 vs. IL-10 + IL-18). (D) Representative confocal laser scanning immunofluorescence images of OPN (red), thrombin (green), and their merge with DAPI (blue) in each Mφ subset. Scale bar represents 20 µm. Higher magnification images are from the white rectangle region in merged panel of Mφ (IL-10 + IL-18). Scale bar represents 10 µm. (E) Relative mean fluorescence intensity (MFI) of CD163 was measured by FACS analysis in each Mφ subset. Hirudin, a specific thrombin inhibitor, was used at 1 µg/mL, n = 3 (***p < 0.001 vs. untreated, ###p < 0.001 vs. IL-10 alone, †††p < 0.001 vs. IL-10 + IL-18). (F) The total areas and lengths of tube-like structures were determined by the Matrigel tube formation assay where b.End5 were cocultured with each Mφ subset. Hirudin was used at 1 µg/mL, n = 6 (***p < 0.001, **p < 0.01 vs. untreated, ##p < 0.01, #p < 0.05 vs. IL-10 alone, †††p < 0.001 vs. IL-10 + IL-18). All data are expressed as means ± SEM and were analyzed by a one-way ANOVA followed by Tukey’s test. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29559970), licensed under a CC-BY license. Not int

Localized differentiation of liver progenitors in arrayed patterns.(A) Immunolabeling of BMEL cells for the biliary marker OPN and hepatocyte marker ALB on arrayed collagen I patterns with control IgG or Fc-recombinant Notch ligands DLL1, DLL4, and JAG1. (B) Quantification of OPN+ cell counts as a function of radial distance from the centroid of each island. (C) Quantification of ALB+ cell counts as a function of radial distance from the centroid of each island. (D) Immunolabeling of BMEL cells presented with DLL4 for the biliary transcription factor SOX9 and hepatocyte transcription factor HNF4A. Arrow in each image indicates the same SOX9+/HNF4A− cell. Scale bar indicates 75 µm. (E, F) Regression analysis of OPN+ and ALB+ cell counts. Data in Figure 1B and Figure 1C were separated into peripheral and central subsets for which dimensionless radius was greater than 0.75 (R>0.75) and less than 0.75 (R<0.75). Separate multiple regression models were generated for each data subset for which coefficient estimates (corresponding to mean change in cell counts) and 95% CI were plotted for OPN+ (E) and ALB+ (F) cells. For each factor, 95% CI that do not intersect with the dashed line indicate regression coefficient estimates for which P<0.05. (A, E) Scale bars indicate 150 µm.10.7554/eLife.38536.007Figure 1—source data 1.Summary table for OPN data in Figure 1B.10.7554/eLife.38536.008Figure 1—source data 2.Summary table for ALB data in Figure 1C.Summary table for OPN data in Figure 1B.Summary table for ALB data in Figure 1C.Immunolabeling and quantification of CK19.(A) Immunolabeling of BMEL cells presented with IgG for the biliary marker CK19. (B) Quantification of CK19 intensity in BMEL cells presented with IgG as a function of radial distance from the centroid of each island. Asterisk (*) indicates P<0.001 for peak intensity (R>0.9) compared with central intensity (R<0.1) using Welch’s t-test.Immunolabeling of OPN and CK19 at t=24h and cell density with radius at t=72h.(A

Aging results in diminished OPN production in response to 2-20 RSV infection.Young and aged BALB/c mice (n = 4/group) were intranasally infected with a dose of 106 pfu/mouse of A2 or 105 pfu/mouse of 2-20) and total lung RNA was collected on time points indicated. (A-B) RNA transcripts of OPN were analyzed with qRT-PCR and represented as a relative ratio of target gene expression to endogenous mouse HPRT and examined with rA2-L19F, 2-20, or A2. (B) 5 µm lung sections obtained at 4, 5 and 8 dpi with either rA2-L19F, 2-20, or A2 infected young and aged mice. Lung sections were immunostained for anti-mouse OPN and nickel-DAB reagent before counterstaining with hematoxylin and eosin. Representative images shown are at 100x magnification with inset [400x] displaying nickel-DAB (dark brown/black staining) positive cells and contrast the hematoxylin (light blue) nuclear stain. Representative images are shown with scale bar indicating 100 µm. (C) Enumeration of OPN-positive cells was performed with ImmunoRatio ImageJ analysis on 200X magnified lung sections from 8 dpi and values are shown as a percentage of total hematoxylin-stained cells in an individual box plot with mean interval bars. Within a single frame, at least 5 frames per mouse (n = 4/group) were collected and individual dot plots are shown of either aged or young mice with interval bars and significance, determined with ANOVA and Fisher's test (p<0.05). (D) qRT-PCR was performed on total lung RNA from young and aged 2–20 RSV infected mice for mRNA expression of OPN receptor CD44. Statistical significance was determine with ANOVA 2-way analysis with p<0.05. All experiments were performed in triplicate. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24558422), licensed under a CC-BY license. Not internally tested by R&D Systems.

Murine placenta cryosections stained with osteopontin (red). Tissue autofluorescence in blue. Image from a verified customer review.

Lkb1 deletion in Ctsk-Cre–expressing cells leads to increased bone formation.(A) μCT analysis of femurs from 5-week-old female Ctsk-Ctrl and Ctsk-CKO mice. (B) Ct.Th and BV/TV of the cortical bones of femurs from 5-week-old female Ctsk-Ctrl and Ctsk-CKO mice (n = 5 for each group). (C) von Kossa staining of the tibiae from 5-week-old female Ctsk-Ctrl and Ctsk-CKO mice. Scale bar: 100 μm. (D and E) Calcein–alizarin red double-labeled fluorescence of the tibiae from 5-week-old female Ctsk-Ctrl and Ctsk-CKO mice showing MAR and BFR of the periosteum (n = 3 for each group). Scale bars: 100 μm (left); 10 μm (right). (F) Gene expression of markers for osteoblast progenitor (Runx2 and Osx), preosteoblast (Alp and Col1a1), and mature osteoblast (Opn, Bsp, and Ocn) of the cortical bones of 20-week-old female Ctsk-CKO mice (n = 3) compared with Ctsk-Ctrl mice (n = 4) tibiae. (G and H) Immunostaining of OSX (G) and OPN (H) in the tumor osteoid of Ctsk-CKO tibiae. Scale bars: 50 μm. Data are represented as mean ± SEM. *P < 0.05; **P < 0.01, unpaired Student’s t test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30830877), licensed under a CC-BY license. Not internally tested by R&D Systems.

OPN expression in the BMF of NOD+/+ mice in the presence of Leishmania amazonensis.(A) Immunofluorescence staining of BMFs. a & d: non-infected control at 24 h and 48 h, respectively. b & c: BMFs infected with LV79 at 24 h and e & f at 48 h post-infection. Green: OPN, red: LV79; blue: nuclei. (B) qRT‒PCR of OPN transcripts (*P = 0.0294). (C) OPN protein quantification in control non-infected (ctl) and infected +LV79 BMFs. NOD ctl 24 h vs NOD+lv79 24 h (*P = 0.0454); NOD ctl 48 h vs NOD+lv79 48 h (**P = 0.0091). Two-tailed Mann‒Whitney test. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0308868), licensed under a CC-BY license. Not internally tested by R&D Systems.

Metformin treatment increases OPN expression in preosteoblasts, contributing to an increase in adhesion of myeloma cells. (A) 2T3, MC3T3 and ST2 were treated with 5 mM metformin for 48 h and osteopontin gene expression quantified (** p < 0.01 as compared to control, n = 3–7). (B) Osteopontin protein expression following metformin treatment for 48 h in 2T3 preosteoblasts and ST2 stromal cells. (C) OPN gene expression in 2T3 preosteoblasts after transfection with either scrambled (SCR) or OPN siRNA (** p < 0.01 as compared to scrambled transfection, n = 3). (D) OPN protein expression in 2T3 preosteoblasts following transfection with OPN siRNA or scrambled control SCR and treatment with metformin. CTR = untransfected. 2T3 preosteoblasts (scrambled control (SCR) or osteopontin siRNA (OPN)) were pretreated with metformin and adhesion imaged and quantified. (E) Representative image and quantification of specific image. (F) Quantification of myeloma cell adhesion (* p < 0.05 as compared to control, n = 4). Data are presented as mean ± SEM. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34890968), licensed under a CC-BY license. Not internally tested by R&D Systems.

Angiogenic potential of retinal microglia during RAP. (A,B) Readplot using the log2 transformation of normalized reads visualizing differentially expressed genes between microglia from Ctrl; MGTom mice (y-axis) and Vldlr−/−; MGTom mice (x-axis) on postnatal day (p)17 (A) and p31 (B). Genes associated with angiogenesis are labeled blue. (C,D) Comparison of RNA Seq results of isolated microglia and protein analysis of retinal tissue lysates from Vldlr−/−; MGTom and Ctrl; MGTom mice at postnatal day (p)17 and p31 for Vegf (C) and Ccl12 (D). mRNA: n = 5–7 samples per group, protein n = 5 samples per group. (E,F) Intravitreal injection of anti-SPP1 ((E), n = 5) or anti-CCL12 ((F), n = 10) in comparison to an IgG injection (*, p < 0.05; **, p < 0.01; ***, p < 0.001). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35408803), licensed under a CC-BY license. Not internally tested by R&D Systems.

Gene and protein expression in muscles from 3m mdx, mdx+ABX, and GFmdxA–KCropped images of representative WB and RT‐qPCR analysis of TA muscle of 3m mdx (n = 3/4), 3m mdx+ABX (n = 3/4), and 3m GFmdx (n = 5) showing the expression of the proteins specifically involved in inflammation/fibrosis (A), skeletal muscle metabolism (B–E), mitochondrial biogenesis (F and G), calcium conducting channels (H and I), autophagy (J), and nicotinic acetylcholine receptors (K). Densitometric data were normalized on vinculin and expressed as mean ± SD. Data are presented as mean ± SD (*P < 0.05, **P < 0.01, ***P < 0.001; ****P < 0.0001, ordinary one‐way ANOVA, Tukey's multiple‐comparison test for WB and non‐parametric test followed by Kruskal–Wallis test for RT‐qPCR).Source data are available online for this figure. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36533294), licensed under a CC-BY license. Not internally tested by R&D Systems.

iOPN promoted STAT1-mediated immunosuppression in MSCs. a–d Vector-MSCs and iOPN-MSCs or WT-MSCs and OPN−/−-MSCs were treated with or without TNF-alpha plus IFN-gamma (10 ng/mL) for the indicated times. Cells were harvested, and OPN, NF-kappa B p65, STAT1, IKB alpha , phosphorylation of NF-kappa B p65, phosphorylation of STAT1 at Tyr701 and phosphorylation of IKB alpha were analyzed by immunoblotting analysis. Full-length blots are presented in Additional file 1: Fig. 5a-d. e–g Vector-MSCs and iOPN-MSCs were stimulated with TNF-alpha plus IFN-gamma (10 ng/mL) for 12 h. The STAT1 inhibitor fludarabine (Flu, 2 μM) was then added to the culture medium of Vector-MSCs and iOPN-MSCs. The expression of iNOS was determined by quantitative real-time PCR (e), immunoblotting analysis (f) and flow cytometry (g). Full-length blots are presented in Additional file 1: Fig. 5f. h Vector-MSCs and iOPN-MSCs were pretreated with DMSO or fludarabine (Flu, 2 μM) for 6 h and then irradiated and cocultured with CFSE-labeled splenocytes activated by anti-CD3/CD28 antibodies for 3 days at a ratio of 1:20. CD4+ T cells and CD8+ T cells were stained for proliferation analysis by flow cytometry at the end of coculture, and the percentages of proliferating T cells are shown. The results are representative of three to six independent experiments. Values are shown as the mean ± SEM and statistical significance is indicated as *P < 0.05, **P < 0.01 and ***P < 0.001 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/39407354), licensed under a CC-BY license. Not internally tested by R&D Systems.

$Lymphatic drainage supports osteoblast survival and BMSC proliferation. A Representative immunofluorescence staining on fracture sides using an anti-OPN antibody to label OBs (green) and an anti-tunel antibody for cell apoptosis (red). Scale bars, 50 um. b Quantitative analysis of OBs apoptosis at fracture sides (n = 5/group, one-way ANOVA). c Scheme of the experimental procedure in vitro for OBs and BMSCs treated by bone marrow or hematoma CM. Bone marrow of sham group and hematoma of fractured group were collected to generate hematoma CM. Rat BMSCs and OBs were cultured with hematoma CM for 24 h and subjected to growth and proliferation analyses. The scheme was modified from Servier Medical Art (//smart.servier.com/), licensed under a Creative Common Attribution 4.0 Generic License (//creativecommons.org/licenses/by/4.0/). d Rat OBs were intervened with hematoma CM for 1 h and OB apoptosis was evaluated by flow cytometry with FITC-Annexin V and PI double staining (n = 5/group, one-way ANOVA). e Rat OBs were cultured with hematoma CM for 1 h and the growth of rat OBs was observed under a microscope. Scale bars, 50 um. f The cell count of rat OBs cultured with hematoma CM for 24 h. g Rat OBs were cultured with hematoma CM for 24 h and the cell proliferation was evaluated using a CCK8 kit. h Rat BMSCs were cultured with hematoma CM for 1 h and the growth of rat BMSCs was observed under a microscope. Scale bars, 50 um. i The cell count of rat BMSCs cultured with hematoma CM for 24 h. j Rat BMSCs, were cultured with hematoma CM for 24 h and the cell proliferation was evaluated using a CCK8 kit. Data are means ± SD. In f,g,i and j, n = 3 wells with biological replicates in the control, sham, VEH and CLO groups, one-way ANOVA. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/39827193), licensed under a CC-BY license. Not internally tested by R&D Systems.$

441-OP) was serially diluted and captured by Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_lightbox">

Recombinant Mouse Osteopontin/OPN (Catalog # <a class=

441-OP) was serially diluted and captured by Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_thumb" />

441-OP) was serially diluted and captured by Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_lightbox">

441-OP) was serially diluted and captured by Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_thumb" />

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF808U-100UG
Alexa Fluor 405	AF808V-100UG
Alexa Fluor 488	AF808G-100UG
Alexa Fluor 532	AF808X-100UG
Alexa Fluor 594	AF808T-100UG
Alexa Fluor 647	AF808R-100UG
Alexa Fluor 700	AF808N-100UG
Alexa Fluor 750	AF808S-100UG
Alexa Fluor Plus 405	AF808AFP405-100UG
Alexa Fluor Plus 488	AF808AFP488-100UG
Alexa Fluor Plus 555	AF808AFP555-100UG
Alexa Fluor Plus 594	AF808AFP594-100UG
Alexa Fluor Plus 647	AF808AFP647-100UG
Alexa Fluor Plus 680	AF808AFP680-100UG
Allophycocyanin	AF808A
Biotin	BAF808
CoraFluor 1	AF808CL1
DyLight 350	AF808D
DyLight 405	AF808E
DyLight 488	AF808K
DyLight 550	AF808L
DyLight 594	AF808M
DyLight 650	AF808W
DyLight 680	AF808Y
DyLight 755	AF808Z
FITC	AF808F
HRP	AF808H
Janelia Fluor 525	AF808JF525
Janelia Fluor 549	AF808I
Janelia Fluor 646	AF808J
Janelia Fluor 669	AF808JF669
PerCP	AF808C
Phycoerythrin	IC808P
mFluor Violet 450 SE	AF808MFV450
mFluor Violet 500 SE	AF808MFV500
mFluor Violet 610 SE	AF808MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC808P
Carrier Free	AF808-SP

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DOST00

Human Osteopontin (OPN) Quantikine ELISA Kit

(80 Publications)

1 Kit

DOST00

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Conjugate	Catalog #	Availability	Size	Price
HRP	PDOST00
HRP	SOST00

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NB110-89062

Osteopontin/OPN Antibody (1B20) - BSA Free

(2 Reviews)

(38 Publications)

0.1 ml

NB110-89062

<b>Biological Strategies Validation. </b>Osteopontin-OPN-Antibody-1B20-BSA-Free-Immunohistochemistry-NB110-89062-img0012.jpg

Osteopontin-OPN-Antibody-1B20-BSA-Free-Immunohistochemistry-NB110-89062-img0013.jpg

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Analysis of Osteopontin on human lung adenocarcinoma.

<b>Biological Strategies Validation. </b>Osteopontin-OPN-Antibody-1B20-BSA-Free-Immunohistochemistry-NB110-89062-img0011.jpg

Simple Western: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Simple Western lane view shows a specific band for Osteopontin in 0.5 mg/ml of Human Breast Cancer lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Photomicrographs of immunostaining for osteocalcin & osteopontin in (A) Group I, (B) Group II (C) Group III at 6 weeks.The pictures are arranged by staining technique (columns) & the investigated treatment (rows). Areas that stained positive for osteocalcin & osteopontin are indicated by red arrowheads. NB, new bone; OS, osteoid; FM, fatty marrow. Image collected & cropped by CiteAb from the following publication (//peerj.com/articles/3513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Photomicrographs of immunostaining for osteocalcin & osteopontin in (A) Group I, (B) Group II (C) Group III at 6 weeks.The pictures are arranged by staining technique (columns) & the investigated treatment (rows). Areas that stained positive for osteocalcin & osteopontin are indicated by red arrowheads. NB, new bone; OS, osteoid; FM, fatty marrow. Image collected & cropped by CiteAb from the following publication (//peerj.com/articles/3513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Osteopontin/OPN (1B20) was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rat anti- Osteopontin/OPN (1B20) Protein G Purified Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB110-89062AF488) (green) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.

Reactivity:

Hu, Mu, Rt, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Rb

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB110-89062AF350
Alexa Fluor 405	NB110-89062AF405
Alexa Fluor 488	NB110-89062AF488
Alexa Fluor 532	NB110-89062AF532
Alexa Fluor 594	NB110-89062AF594
Alexa Fluor 647	NB110-89062AF647
Alexa Fluor 700	NB110-89062AF700
Alexa Fluor 750	NB110-89062AF750
Alexa Fluor Plus 405	NB110-89062AFP405
Alexa Fluor Plus 488	NB110-89062AFP488
Alexa Fluor Plus 555	NB110-89062AFP555
Alexa Fluor Plus 594	NB110-89062AFP594
Alexa Fluor Plus 647	NB110-89062AFP647
Alexa Fluor Plus 680	NB110-89062AFP680
Allophycocyanin	NB110-89062APC
Biotin	NB110-89062B
CoraFluor 1	NB110-89062CL1
DyLight 350	NB110-89062UV
DyLight 405	NB110-89062V
DyLight 488	NB110-89062G
DyLight 550	NB110-89062R
DyLight 594	NB110-89062DL594
DyLight 650	NB110-89062C
DyLight 680	NB110-89062FR
DyLight 755	NB110-89062IR
FITC	NB110-89062F
HRP	NB110-89062H
Janelia Fluor 525	NB110-89062JF525
Janelia Fluor 549	NB110-89062JF549
Janelia Fluor 646	NB110-89062JF646
Janelia Fluor 669	NB110-89062JF669
PE	NB110-89062PE
PerCP	NB110-89062PCP
mFluor Violet 450 SE	NB110-89062MFV450
mFluor Violet 500 SE	NB110-89062MFV500
mFluor Violet 610 SE	NB110-89062MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB110-89062SS
Azide and BSA Free	NBP2-80895

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AF1433

Osteopontin/OPN Antibody [Unconjugated]

(51 Publications)

100 μg

AF1433

NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox">

Osteopontin/OPN was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line (positive staining; left panel) and Daudi human Burkitt's lymphoma cell line (negative staining; right panel) using Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # <a class=

NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." alt="Osteopontin/OPN was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line (positive staining; left panel) and Daudi human Burkitt's lymphoma cell line (negative staining; right panel) using Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the surface of epithelial cells in the intralobular duct. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_lightbox">

Osteopontin/OPN was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using 8 µg/mL Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # <a class=

CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the surface of epithelial cells in the intralobular duct. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_thumb" />

CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_lightbox">

Osteopontin/OPN was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # <a class=

CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_thumb" />

1433-OP), immobilized onto a microplate, supports the adhesion of the HEK293 human embryonic kidney cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human Osteopontin/OPN (1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433). The ND₅₀ is typically 0.300 - 3.60 µg/mL." class="big_lightbox">

Recombinant Human Osteopontin/OPN (Catalog # <a class=

1433-OP), immobilized onto a microplate, supports the adhesion of the HEK293 human embryonic kidney cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human Osteopontin/OPN (1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433). The ND₅₀ is typically 0.300 - 3.60 µg/mL." class="big_thumb" />

MAB14332R) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994)." class="big_lightbox">

Recombinant Human Osteopontin/OPN protein was serially diluted 2-fold and captured by Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # <a class=

MAB14332R) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994)." class="big_thumb" />

AF1433) at 1ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in tubules. " class="big_lightbox">

Formalin-fixed paraffin-embedded tissue sections of human kidney were probed for OPN mRNA (ACD RNAScope Probe, catalog #420101; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-human OPN polyclonal antibody (R&D Systems catalog # <a class=

AF1433) at 1ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in tubules. " class="big_thumb" />

DKK1 blockade inhibits mineralization of osteoblast differentiation. Osteoblasts were treated with 1 μg anti-DKK1 or IgG as controls during osteoblast differentiation (n = 3). At indicated days, analysis of (A) results of ALP and ARS staining; scale bar is 200 μm, (B) hydroxyapatite staining; BF, Bright Field; HA, Hydroxyapatite; Scale bar is 500 μm, (C) immunoblotting for proteins, and (D) qPCR for mRNA. Representative data are shown (n = 3). * p < 0.05 (mean ± SD; n = 3). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31963554), licensed under a CC-BY license. Not internally tested by R&D Systems.

Molecular mechanism underlying CAF‐derived SPP1‐induced resistance to sorafenib or lenvatinib. (A‐B) Changes in the expression levels of PKC alpha , BRAF/ERK/STAT3, and PI3K/AKT/mTOR pathway proteins in CAF‐CM‐incubated Huh‐7 cells treated with sorafenib (15 μmol/L; A) or lenvatinib alone (5 μmol/L; B) and in combination with SPP1‐BP. (C‐D) Changes in the phosphorylation of AKT, mTOR, BRAF, and ERK1/2 under ITGB1 and/or ITGB5 silencing in Hep3B and Huh‐7 cells treated with sorafenib (C) or lenvatinib (D) in combination with CAF‐CM. (Mean ± SEM; Two‐way ANOVA test; *P < 0.05; **P < 0.01; ***P < 0.001).Abbreviations: DMSO, dimethyl sulfoxide; CAF, cancer‐associated fibroblast; CM, culture medium; Sor, sorafenib; SPP1‐BP, SPP1‐blocking peptide, PKC alpha , protein kinase C alpha ; BRAF,v‐Raf murine sarcoma viral oncogene homolog B; ERK, extracellular signal‐related kinase; STAT3, signal transducer and activator of transcription 3; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; PI3K, phosphatidylinositol‐3‐kinase; AKT, protein kinase B; mTOR, mammalian target of rapamycin; Len, lenvatinib; siCtrl, negative control; ITGB1, integrin subunit beta 1; ITGB5, integrin subunit beta 5. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36919193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Identification of the CAF‐derived molecules that induce resistance to sorafenib or lenvatinib in patients with HCC. (A) Heat map of 790 genes overexpressed in CAFs. (B) GSEA of the genes overexpressed in CAFs. (C) Process of candidate gene selection. (D) Expression levels of the 9 candidate genes according to sorafenib response in the GSE109211 dataset. (E) Comparison of SPP1 expression between sorafenib responders and non‐responders in the GSE109211 and GSE143233 datasets. (F) Immunofluorescence staining of alpha ‐SMA and SPP1 in tumor tissues from patients with HCC. (G) Comparison of SPP1 expression between CAFs and their paired para‐cancer fibroblasts in the WTS data from the 9 pairs of CAF and para‐cancer fibroblasts. (H) Representative H&E and SPP1 IHC images of the tumor (T), tumor stroma (S) (left), and non‐tumor fibrous tissue (middle) in an SPP1‐high patient. Scale bar, 50 μm. Comparison of SPP1 IHC staining intensity according to tumor location in the SPP1‐high group (right). (I) Representative H&E and SPP1 IHC images of tumor (T), tumor stroma (S) (left), and non‐tumor fibrous tissue (middle) in an SPP1‐low patient. Comparison of SPP1 IHC staining intensity according to tumor location in the SPP1‐low group (right). (Mean ± SEM; unpaired Welch's t‐test; *P < 0.05; **P < 0.01; ***P < 0.001).Abbreviations: HCC, hepatocellular carcinoma; NF, normal fibroblast; CAFs, cancer‐associated fibroblasts; ELISA, enzyme linked immunosorbent assay; H&E, hematoxylin and eosin; alpha ‐SMA, alpha smooth muscle actin; SPP1, secreted phosphoprotein 1; DAPI, 4′, 6‐diamidino‐2‐phenylindole; PAF, para‐cancer fibroblast; S, tumor stroma; T, tumor; IHC, immunohistochemistry. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36919193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Integrin complexes (integrin alpha V beta 5, alpha 5 beta 1, and alpha V beta 1) on HCC cells were identified as binding proteins of SPP1. (A) Western blotting of SPP1 receptor‐binding protein expression in HCC cells. (B) Measurement of CD44 and integrin expression in Hep3B and Huh‐7 cells by fluorescent intensity. (C‐E) Co‐IP of SPP1 after CAF‐CM treatment, followed by Western blotting for SPP1, ITGA5, ITGAV, ITGB1, and ITGB5 in HCC cells (C), HCC cells silenced for ITGB1 and/or ITGB5 (D), and HCC cells incubated with/without SPP1‐BP (E).Abbreviations: ITGA4, integrin subunit alpha 4; ITGA5, integrin subunit alpha 5; ITGA8, integrin subunit alpha 8; ITGA9, integrin subunit alpha 9; ITGAV, integrin subunit alpha V; ITGB1, integrin subunit beta 1; ITGB3, integrin subunit beta 3; ITGB5, integrin subunit beta ; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence units; IgG, immunoglobulin G; SPP1, secreted phosphoprotein 1; CAF, cancer‐associated fibroblast; CM, culture medium; IB, immunoblotting; siCtrl, negative control; SPP1‐BP, SPP1‐blocking peptide. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36919193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Evaluation of SPP1 expression on cancer cells and macrophages. (A,B). Representative dual ISH (green: SPP1; red: Iba-1) in adenocarcinoma. (C). Representative single-IHC of SPP1 in adenocarcinoma and SCC. (D). Double-IHC of SPP1 and Iba-1 to examine the SPP1 expression level in cancer cells (Iba-1-negative). Double-IHC of SPP1 and PU.1 to examine the SPP1 expression level on TAMs (PU.1-positive in the nucleus). (E). SPP1’s expression level was scored according to the proportion of stained cells as follows: less than 1% staining, score 0; 1% to 49% staining, score 1; more than 50% staining, score 2. The proportions of samples with each score among cancer cells and TAMs in adenocarcinoma and SCC are shown in the pie charts. Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/2072-6694/14/18/4374), licensed under a CC-BY license. Not internally tested by R&D Systems.

SPP1 expression in macrophages. (A) SPP1 expression in macrophages, THP-1 cells, U937 cells, and 10 lung cancer cell lines was examined by qRT-PCR. (B) Macrophages (blue) and THP-1 cells (green) were stimulated with the CM of cancer cell lines; then, the SPP1 mRNA expression level was evaluated. Control (CT) refers to macrophages or THP-1 without CM of cancer cell lines added. (C) Monocytes were cultured with M-CSF and GM-CSF as described in the Materials and Methods section, and SPP1 mRNA and CD204 mRNA levels were evaluated at different stages of monocyte/macrophage culture (days 0, 3, and 5). (D) Monocytes were cultured with M-CSF or GM-CSF for 5 days, and SPP1 and CD204 expression levels were examined. (E) SPP1′s concentration (ng/mL) in the CM of macrophages cultured with M-CSF or GM-CSF was examined by ELISA at days 1 to 3 (n = 3). (F) Immunocytostaining of SPP1 in the macrophages cultured with M-CSF or GM-CSF at days 0 and 5. (G) SPP1 and CD204 expression levels in THP-1 cells differentiated for 0, 1, 3, or 6 days were examined by qRT-PCR (G) and Western blot analysis (H). *: statistically significant, p value < 0.05. Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/2072-6694/14/18/4374), licensed under a CC-BY license. Not internally tested by R&D Systems.

1433-OP) was serially diluted and captured by Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14332) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_lightbox">

1433-OP) was serially diluted and captured by Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14332) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_thumb" />

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF1433U-100UG
Alexa Fluor 405	AF1433V-100UG
Alexa Fluor 488	AF1433G-100UG
Alexa Fluor 532	AF1433X-100UG
Alexa Fluor 594	AF1433T-100UG
Alexa Fluor 647	AF1433R-100UG
Alexa Fluor 700	AF1433N-100UG
Alexa Fluor 750	AF1433S-100UG
Alexa Fluor Plus 405	AF1433AFP405-100UG
Alexa Fluor Plus 488	AF1433AFP488-100UG
Alexa Fluor Plus 555	AF1433AFP555-100UG
Alexa Fluor Plus 594	AF1433AFP594-100UG
Alexa Fluor Plus 647	AF1433AFP647-100UG
Alexa Fluor Plus 680	AF1433AFP680-100UG
Allophycocyanin	AF1433A
Biotin	BAF1433
CoraFluor 1	AF1433CL1
DyLight 350	AF1433D
DyLight 405	AF1433E
DyLight 488	AF1433K
DyLight 550	AF1433L
DyLight 594	AF1433M
DyLight 650	AF1433W
DyLight 680	AF1433Y
DyLight 755	AF1433Z
FITC	AF1433F
HRP	AF1433H
Janelia Fluor 525	AF1433JF525
Janelia Fluor 549	AF1433I
Janelia Fluor 646	AF1433J
Janelia Fluor 669	AF1433JF669
PE	AF1433P
PerCP	AF1433C
mFluor Violet 450 SE	AF1433MFV450
mFluor Violet 500 SE	AF1433MFV500
mFluor Violet 610 SE	AF1433MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF1433-SP

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MOST00

Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit

(46 Publications)

1 Kit

MOST00

Reactivity:

Mu, Rt

Species Glossary

Reactivity: Mu, Rt

Applications:

Applications:

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1433-OP

Recombinant Human Osteopontin (OPN) Protein

(47 Publications)

50 μg

1433-OP

1 μg/lane of Recombinant Human Osteopontin/OPN was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a bands at 60-65 kDa.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	1433-OP-050/CF

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441-OP

Recombinant Mouse Osteopontin/OPN Protein

(44 Publications)

50 μg

441-OP

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

Applications:

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	441-OP-050/CF
Carrier Free	441-OP-200/CF

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DY1433

Human Osteopontin (OPN) DuoSet ELISA, 15 Plate

(33 Publications)

1 Kit (for 15 Plates)

DY1433

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

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DY441

Mouse Osteopontin DuoSet ELISA, 15 Plate

(18 Publications)

1 Kit (for 15 Plates)

DY441

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

Applications:

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MAB1433

Osteopontin/OPN Antibody (190312) [Unconjugated]

(1 Review)

(7 Publications)

100 μg

MAB1433

CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_lightbox">

Osteopontin/OPN was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB1433) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # <a class=

CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." alt="Osteopontin/OPN was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB1433) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_thumb" />

SPP1 and OPN expression in BAL cells. (A) SSP1 relative mRNA levels in IPF and N-IPF relative to healthy (Brown-Forsythe Anova, with individual comparisons), (B) Violin plots of mean OPN expression/cell per disease group (Kruskal-Wallis test and individual comparisons) and (C) Typical Images of BAL cytospins stained with anti-human OPN antibody and ToPro-633 nuclear stain in (i) healthy, (ii) IPF and (iii) N-IPF samples (*p < 0.05, **P < 0.01, ****p < 0.0001). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34867934), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	FAB1433U-100UG
Alexa Fluor 405	FAB1433V-100UG
Alexa Fluor 488	FAB1433G-100UG
Alexa Fluor 532	FAB1433X-100UG
Alexa Fluor 594	FAB1433T-100UG
Alexa Fluor 647	FAB1433R-100UG
Alexa Fluor 700	FAB1433N-100UG
Alexa Fluor 750	FAB1433S-100UG
Alexa Fluor Plus 405	FAB1433AFP405-100UG
Alexa Fluor Plus 488	FAB1433AFP488-100UG
Alexa Fluor Plus 555	FAB1433AFP555-100UG
Alexa Fluor Plus 594	FAB1433AFP594-100UG
Alexa Fluor Plus 647	FAB1433AFP647-100UG
Alexa Fluor Plus 680	FAB1433AFP680-100UG
Allophycocyanin	FAB1433A
Biotin	FAB1433B
CoraFluor 1	FAB1433CL1
DyLight 350	FAB1433D
DyLight 405	FAB1433E
DyLight 488	FAB1433K
DyLight 550	FAB1433L
DyLight 594	FAB1433M
DyLight 650	FAB1433W
DyLight 680	FAB1433Y
DyLight 755	FAB1433Z
FITC	FAB1433F
HRP	FAB1433H
Janelia Fluor 525	FAB1433JF525
Janelia Fluor 549	FAB1433I
Janelia Fluor 646	FAB1433J
Janelia Fluor 669	FAB1433JF669
PE	FAB1433P
PerCP	FAB1433C
mFluor Violet 450 SE	FAB1433MFV450
mFluor Violet 500 SE	FAB1433MFV500
mFluor Violet 610 SE	FAB1433MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB1433-SP

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NB600-1043

Osteopontin/OPN Antibody - BSA Free

(8 Publications)

0.1 ml

NB600-1043

Rabbit anti-Osteopontin was used at a 1:100-1:300 dilution to detect osteopontin by immunohistochemistry. Osteopontin is a normal component of elastic fibers in the breast (shown heavily stained in this section of human breast tumor cells). There is also weak staining of the extracellular matrix. Osteopontin is not expressed in breast tumor cells, and there is no staining of the breast cells in this section. No antigen retrieval is required.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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MAB14331

Osteopontin/OPN Antibody (223126) [Unconjugated]

(9 Publications)

100 μg

MAB14331

HAF007). A specific band was detected for Osteopontin/OPN at approximately 60 to 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

Western blot shows lysates of human adipose (diabetes) tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14331) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # <a class=

HAF007). A specific band was detected for Osteopontin/OPN at approximately 60 to 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of human adipose (diabetes) tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14331) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Osteopontin/OPN at approximately 60 to 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

Simple Western lane view shows lysates of human adipose tissue and human adipose (diabetes) tissue, loaded at 0.2 mg/mL. A specific band was detected for Osteopontin/OPN at approximately 63 kDa (as indicated) using 100 µg/mL of Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14331). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.</PP>*Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

MAB003, open histogram) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules." class="big_lightbox">

U937 cells were stained with Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # mab14331, filled histogram) or isotype control antibody (Catalog # <a class=

MAB003, open histogram) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules." class="big_thumb" />

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

,

Applications: ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	FAB14331U-100UG
Alexa Fluor 405	FAB14331V-100UG
Alexa Fluor 488	FAB14331G-100UG
Alexa Fluor 532	FAB14331X-100UG
Alexa Fluor 594	FAB14331T-100UG
Alexa Fluor 647	FAB14331R-100UG
Alexa Fluor 700	FAB14331N-100UG
Alexa Fluor 750	FAB14331S-100UG
Alexa Fluor Plus 405	FAB14331AFP405-100UG
Alexa Fluor Plus 488	FAB14331AFP488-100UG
Alexa Fluor Plus 555	FAB14331AFP555-100UG
Alexa Fluor Plus 594	FAB14331AFP594-100UG
Alexa Fluor Plus 647	FAB14331AFP647-100UG
Alexa Fluor Plus 680	FAB14331AFP680-100UG
Allophycocyanin	IC14331A
Biotin	FAB14331B
CoraFluor 1	FAB14331CL1
DyLight 350	FAB14331D
DyLight 405	FAB14331E
DyLight 488	FAB14331K
DyLight 550	FAB14331L
DyLight 594	FAB14331M
DyLight 650	FAB14331W
DyLight 680	FAB14331Y
DyLight 755	FAB14331Z
Fluorescein	IC14331F
HRP	FAB14331H
Janelia Fluor 525	FAB14331JF525
Janelia Fluor 549	FAB14331I
Janelia Fluor 646	FAB14331J
Janelia Fluor 669	FAB14331JF669
PerCP	FAB14331C
Phycoerythrin	IC14331P
mFluor Violet 450 SE	FAB14331MFV450
mFluor Violet 500 SE	FAB14331MFV500
mFluor Violet 610 SE	FAB14331MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC14331P
Carrier Free	IC14331A
Carrier Free	IC14331F
Carrier Free	MAB14331-SP
Carrier Free	MAB14331-500

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

Modification

Clonality

Clone

Gene Symbol

Images Type

Isotype

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

Promotional Item

Species Glossary

SPP1 Products By Gene Name

Osteopontin/OPN Antibody [Unconjugated]

Human Osteopontin (OPN) Quantikine ELISA Kit

Osteopontin/OPN Antibody (1B20) - BSA Free

Osteopontin/OPN Antibody [Unconjugated]

Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit

Recombinant Human Osteopontin (OPN) Protein

Recombinant Mouse Osteopontin/OPN Protein

Human Osteopontin (OPN) DuoSet ELISA, 15 Plate

Mouse Osteopontin DuoSet ELISA, 15 Plate

Osteopontin/OPN Antibody (190312) [Unconjugated]

Osteopontin/OPN Antibody - BSA Free

Osteopontin/OPN Antibody (223126) [Unconjugated]