S100A8 Antibody [Unconjugated]

Western blot shows lysates of mouse spleen tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse S100A8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3059) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for S100A8 at approximately 10-11 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

S100A8 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse S100A8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3059) at 10 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 counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

S100A8 was detected in immersion fixed NMuMG mouse mammary gland epithelial cell line using 10 µg/mL Goat Anti-Mouse S100A8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3059) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western lane view shows lysates of mouse spleen tissue, loaded at 0.2 mg/mL. A specific band was detected for S100A8 at approximately 9 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Mouse S100A8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3059) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.

S100A8/A9 proteins augment differentiation and osteoclastic activity derived from iOCPs but not from hOCPs. a Expression of S100A8, S100A9 and RAGE in sorted control and inflamed BM iOCPs and hOCPs. The protein phosphatase 2A catalytic subunit (PP2Ac) is shown as a loading control. b Densitometry measurements of three biological repeats (no statistical analysis presented). c Sorted control and inflamed BM iOCPs and hOCPs (5 × 104) were cultured on the Osteo assay surface with or without anti-RAGE blocking antibodies (bar: 50 µm). d Pit area quantitation. e Sorted iOCPs and hOCPs (5 × 104) from the BM of control mice were cultured on the Osteo assay surface in combination with a recombinant S100A8/A9 heterodimer and anti-RAGE blocking antibodies as indicated (bar: 50 µm). f Pit area quantitation. c–f Depict representative results for two independent experiments, n = 5 for each group. Line: median, box: 25th-75th percentile, whiskers: range. *P < 0.05 (Mann–Whitney test and Holm multiplicity correction) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35396510), licensed under a CC-BY license. Not internally tested by R&D Systems.

iASPP deficient keratinocytes induce pro-inflammatory gene expression in vitro and attract macrophages in vivo(A) qPCR analysis of mRNA expression levels of inflammatory genes (and iASPP gene Ppp1r13l) in iASPP WT and KO primary keratinocytes at 0, 1, and 6 h after treatment with TNF-alpha . Values are mean + SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; n = 3 biological replicates.(B) Left, IHC staining of S100a8 and S100a9 in untreated skin sections from iASPP WT and KO mice (adjacent sections). Scale bar, 50 μm. Right, IB of iASPP and S100a9 expression levels in iASPP WT (−tamoxifen) and KO (+tamoxifen) primary keratinocytes.(C) H&E analysis of acetone- or TPA-treated skin sections from iASPP WT and KO mice. Scale bar, 50 μm. Histograms below show epidermal thickness in the same samples. Values are mean ± SD. n = 3 (WT) and n = 4 (KO) mice in acetone cohort; n = 4 (WT), n = 3 (KO) mice in TPA cohort.(D) IHC staining of F4/80-positive macrophages in acetone- or TPA-treated skin sections from iASPP WT and KO mice. Scale bar, 50 μm. Histograms below show quantification of the same samples. Values are mean + SD. Same cohort as in (C).See STAR Methods for p calculations for (A), (C), and (D). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36261000), licensed under a CC-BY license. Not internally tested by R&D Systems.

iASPP deficient keratinocytes induce pro-inflammatory gene expression in vitro and attract macrophages in vivo(A) qPCR analysis of mRNA expression levels of inflammatory genes (and iASPP gene Ppp1r13l) in iASPP WT and KO primary keratinocytes at 0, 1, and 6 h after treatment with TNF-alpha . Values are mean + SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; n = 3 biological replicates.(B) Left, IHC staining of S100a8 and S100a9 in untreated skin sections from iASPP WT and KO mice (adjacent sections). Scale bar, 50 μm. Right, IB of iASPP and S100a9 expression levels in iASPP WT (−tamoxifen) and KO (+tamoxifen) primary keratinocytes.(C) H&E analysis of acetone- or TPA-treated skin sections from iASPP WT and KO mice. Scale bar, 50 μm. Histograms below show epidermal thickness in the same samples. Values are mean ± SD. n = 3 (WT) and n = 4 (KO) mice in acetone cohort; n = 4 (WT), n = 3 (KO) mice in TPA cohort.(D) IHC staining of F4/80-positive macrophages in acetone- or TPA-treated skin sections from iASPP WT and KO mice. Scale bar, 50 μm. Histograms below show quantification of the same samples. Values are mean + SD. Same cohort as in (C).See STAR Methods for p calculations for (A), (C), and (D). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36261000), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB, Simple Western, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

S100A8 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant mouse S100A8
  Pro2-Glu89
  Accession # P27005
• Specificity: Detects mouse S100A8 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 5% cross-reactivity with recombinant mouse (rm) S100A9, rmS100A10, and recombinant human S100B is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: S100A8
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Simple Western 2.5 ug/mL
  • Western Blot 0.25 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for S100A8 Antibody [Unconjugated]

• 60B8AG
• CAGACP-10
• Calgranulin A
• calgranulin-A
• Calprotectin L1L subunit
• CFAG
• CFAGL1Ag
• CGLA
• Cystic fibrosis antigen
• Leukocyte L1 complex light chain
• MA387
• MIF
• Migration inhibitory factor-related protein 8
• MRP8
• MRP-8
• MRP8S100 calcium binding protein A8 (calgranulin A)
• NIF
• P8
• protein S100-A8
• S100 calcium binding protein A8
• S100 calcium-binding protein A8 (calgranulin A)
• S100 calcium-binding protein A8
• S100A8
• Urinary stone protein band A

Background

Mouse S100A8, also known as CP-10, Calgranulin A and MRP8, is a 10 kDa member of the S100 family of calcium-binding proteins. S100A8 contains short sequential modules, including an N-terminal alpha -helix, a Ca⁺⁺-binding EF-hand segment, a short central linker region, a second EF-hand segment, and a C-terminal alpha -helix. S100A8 noncovalently heterodimerizes with S100A9. In the presence of Ca⁺⁺, the heterodimers form heterotetramers. The dimeric complex is found both intracellularly and extracellularly. It binds to heparan sulfate and is chemotactic for PMNs and macrophages. The amino acid sequence of mouse S100A8 is 80% and 57% identical to that of rat and human S100A8, respectively.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for S100A8 Antibody (AF3059)(27)

Publications using AF3059

• Hashimoto, H;Okazaki, T;Honkura, Y;Ren, Y;Ngamsnae, P;Hisaoka, T;Koshiba, Y;Suzuki, J;Ebihara, S;Katori, Y; Nrf2 Deficiency Exacerbates the Decline in Swallowing and Respiratory Muscle Mass and Function in Mice with Aspiration Pneumonia International journal of molecular sciences 2024-11-04 [PMID: 39519380] (Immunohistochemistry, Transgenic Mouse)
• Yang, Y;Abbas, S;Sayem, MA;Dutta, A;Mohi, G; SRSF2 mutation reduces polycythemia and impairs hematopoietic progenitor functions in JAK2V617F-driven myeloproliferative neoplasm Blood cancer journal 2023-11-27 [PMID: 38012156] (Western Blot, Mouse)
• Wang J, Rattner A, Nathans J Bacterial meningitis in the early postnatal mouse studied at single-cell resolution eLife 2023-06-15 [PMID: 37318981]
• Xu XJ, Ge QQ, Yang MS et al. Neutrophil-derived interleukin-17A participates in neuroinflammation induced by traumatic brain injury Neural regeneration research 2023-05-01 [PMID: 36254991]
• C Conrad, D Yildiz, SJ Cleary, A Margraf, L Cook, U Schlomann, B Panaretou, JL Bowser, H Karmouty-Q, J Li, NK Berg, SC Martin, A Aljohmani, SF Moussavi-H, KM Wang, JJ Tian, M Magnen, C Valet, L Qiu, JP Singer, HK Eltzschig, CAPSys Stu, W Bertrams, S Herold, N Suttorp, B Schmeck, ZT Ball, A Zarbock, MR Looney, JW Bartsch ADAM8 signaling drives neutrophil migration and ARDS severity JCI Insight, 2022-02-08;7(3):. 2022-02-08 [PMID: 35132956] (IHC, Mouse)
• G Theocharid, BE Thomas, D Sarkar, HL Mumme, WJR Pilcher, B Dwivedi, T Sandoval-S, RF Sîrbulescu, A Kafanas, I Mezghani, P Wang, A Lobao, IS Vlachos, B Dash, HC Hsia, V Horsley, SS Bhasin, A Veves, M Bhasin Single cell transcriptomic landscape of diabetic foot ulcers Nature Communications, 2022-01-10;13(1):181. 2022-01-10 [PMID: 35013299] (IHC, Human)
• YT Ho, T Shimbo, E Wijaya, T Kitayama, S Takaki, K Ikegami, K Miyashita, Y Ouchi, E Takaki, R Yamamoto, Y Kaneda, K Tamai Longitudinal single cell transcriptomics reveals a role for Serpina3n-mediated resolution of inflammation in a mouse colitis model Cellular and Molecular Gastroenterology and Hepatology, 2021-04-13;0(0):. 2021-04-13 [PMID: 33862275] (IHC, Mouse)
• Darrell Pilling, Tejas R. Karhadkar, Richard H. Gomer A CD209 ligand and a sialidase inhibitor differentially modulate adipose tissue and liver macrophage populations and steatosis in mice on the Methionine and Choline-Deficient (MCD) diet PLOS ONE 2020-12-30 [PMID: 33378413]
• MJ Kim, JY Kim, M Kang, MH Won, SH Hong, Y Her Reduced Fecal Calprotectin and Inflammation in a Murine Model of Atopic Dermatitis Following Probiotic Treatment Int J Mol Sci, 2020-05-31;21(11):. 2020-05-31 [PMID: 32486523] (Western Blot, Mouse)
• S Maekawa, S Onizuka, S Katagiri, M Hatasa, Y Ohsugi, N Sasaki, K Watanabe, A Ohtsu, R Komazaki, K Ogura, T Miyoshi-Ak, T Iwata, H Nitta, Y Izumi RNA sequencing for ligature induced periodontitis in mice revealed important role of S100A8 and S100A9 for periodontal destruction Sci Rep, 2019-10-11;9(1):14663. 2019-10-11 [PMID: 31605018] (IHC-P, Mouse)
• YE Kim, M Lee, H Gu, J Kim, S Jeong, S Yeo, YJ Lee, SH Im, YC Sung, HJ Kim, IL Weissman, GO Ahn Hypoxia-inducible factor-1 (HIF-1) activation in myeloid cells accelerates DSS-induced colitis progression in mice Dis Model Mech, 2018-07-30;0(0):. 2018-07-30 [PMID: 29967068] (IHC, Western Blot, Mouse)
• A Dufour, CL Bellac, U Eckhard, N Solis, T Klein, R Kappelhoff, N Fortelny, P Jobin, J Rozmus, J Mark, P Pavlidis, V Dive, SJ Barbour, CM Overall C-terminal truncation of IFN-? inhibits proinflammatory macrophage responses and is deficient in autoimmune disease Nat Commun, 2018-06-20;9(1):2416. 2018-06-20 [PMID: 29925830] (Western Blot, Human)
• Exploring experimental cerebral malaria pathogenesis through the characterisation of host-derived plasma microparticle protein content Sci Rep, 2016-12-05;6(0):37871. 2016-12-05 [PMID: 27917875] (Western Blot, Mouse)
• Beneficial Effects of Sodium Phenylbutyrate Administration During Infection with Salmonella enterica serovar Typhimurium Infect Immun, 2016-08-19;0(0):. 2016-08-19 [PMID: 27382022] (Western Blot, Mouse)
• Heather L. Clark, Anupam Jhingran, Yan Sun, Chairut Vareechon, Steven de Jesus de Jesus Carrion, Eric P. Skaar et al. Zinc and Manganese Chelation by Neutrophil S100A8/A9 (Calprotectin) Limits Extracellular Aspergillus fumigatus Hyphal Growth and Corneal Infection The Journal of Immunology 2016-01-01 [PMID: 26582948]
• Thomsen M, Bakiri L, Hasenfuss S, Wu H, Morente M, Wagner E Loss of JUNB/AP-1 promotes invasive prostate cancer. Cell Death Differ, 2014-12-19;22(4):574-82. 2014-12-19 [PMID: 25526087] (Western Blot, Mouse)
• Yajima-Himuro , Sara, Oshima , Masamits, Yamamoto , Gou, Ogawa , Miho, Furuya , Madoka, Tanaka , Junichi, Nishii , Kousuke, Mishima , Kenji, Tachikawa , Tetsuhik, Tsuji , Takashi, Yamamoto , Matsuo The junctional epithelium originates from the odontogenic epithelium of an erupted tooth. Sci Rep, 2014-05-02;4(0):4867. 2014-05-02 [PMID: 24785116] (IHC, Mouse)
• Baluk P, Phillips K, Yao L, Adams A, Nitschke M, McDonald D Neutrophil dependence of vascular remodeling after Mycoplasma infection of mouse airways. Am J Pathol, 2014-04-13;184(6):1877-89. 2014-04-13 [PMID: 24726646] (IHC, Mouse)
• Baluk P, Adams A, Phillips K, Feng J, Hong Y, Brown M, McDonald D Preferential lymphatic growth in bronchus-associated lymphoid tissue in sustained lung inflammation. Am J Pathol, 2014-03-11;184(5):1577-92. 2014-03-11 [PMID: 24631179] (IHC, Mouse)
• Kim H, Chang K, Ha T, Kim J, Ha S, Shin K, Moon C, Nacken W, Kim H, Suh Y S100A9 knockout decreases the memory impairment and neuropathology in crossbreed mice of Tg2576 and S100A9 knockout mice model. PLoS ONE, 2014-02-25;9(2):e88924. 2014-02-25 [PMID: 24586443] (Western Blot, Mouse)
• Behnsen J, Jellbauer S, Wong C, Edwards R, George M, Ouyang W, Raffatellu M The cytokine IL-22 promotes pathogen colonization by suppressing related commensal bacteria. Immunity, 2014-02-06;40(2):262-73. 2014-02-06 [PMID: 24508234] (Western Blot, Mouse)
• Yano J, Palmer G, Eberle K, Peters B, Vogl T, McKenzie A, Fidel P Vaginal epithelial cell-derived S100 alarmins induced by Candida albicans via pattern recognition receptor interactions are sufficient but not necessary for the acute neutrophil response during experimental vaginal candidiasis. 2013-12-09;0(0):. 2013-12-09 [PMID: 24478092] (Neutralization, Mouse)
• Sade-Feldman M, Kanterman J, Ish-Shalom E, Elnekave M, Horwitz E, Baniyash M Tumor necrosis factor-alpha blocks differentiation and enhances suppressive activity of immature myeloid cells during chronic inflammation. Immunity, 2013-03-07;38(3):541-54. 2013-03-07 [PMID: 23477736] (Neutralization, Western Blot, Mouse)
• Corinne Cayatte, Barbara Joyce-Shaikh, Felix Vega, Katia Boniface, Jeffrey Grein, Erin Murphy et al. Biomarkers of Therapeutic Response in the IL-23 Pathway in Inflammatory Bowel Disease Clinical and Translational Gastroenterology 2012-02-16 [PMID: 23238132]
• Yano , Junko, Kolls , Jay K, Happel , Kyle I, Wormley , Floyd, Wozniak , Karen L, Fidel , Paul L J The Acute Neutrophil Response Mediated by S100 Alarmins during Vaginal Candida Infections Is Independent of the Th17-Pathway. PLoS ONE, 2012-09-25;7(9):e46311. 2012-09-25 [PMID: 23050010] (ELISA Development, Mouse)
• Gonzalez-Lopez A, Aguirre A, Lopez-Alonso I, Amado L, Astudillo A, Fernandez-Garcia MS, Suarez MF, Batalla-Solis E, Colado E, Albaiceta GM MMP-8 Deficiency Increases TLR/RAGE Ligands S100A8 and S100A9 and Exacerbates Lung Inflammation during Endotoxemia. PLoS ONE, 2012-06-29;7(6):e39940. 2012-06-29 [PMID: 22768176] (Western Blot, Mouse)
• Cuervo H, Guerrero NA, Carbajosa S, Beschin A, De Baetselier P, Girones N, Fresno M Myeloid-Derived Suppressor Cells Infiltrate the Heart in Acute Trypanosoma cruzi Infection. J. Immunol., 2011-07-29;187(5):2656-65. 2011-07-29 [PMID: 21804013] (Western Blot, Mouse)

Reviews for S100A8 Antibody (AF3059) (2)

Average Rating: 3.5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Mouse.

reviewed by: Verified Customer WB Mouse 01/07/2015

Summary

• Application: Western Blot
• Sample Tested: See PMID 20098622
• Species: Mouse

reviewed by: Verified Customer WB Mouse 01/07/2015

Summary

• Application: Western Blot
• Sample Tested: See PMID 23548910
• Species: Mouse

FAQs for S100A8 Antibody (AF3059). (Showing 1 - 2 of 2 FAQs).

1. Have any of your MRP8 antibodies been tested for use in neutralization, or blocking assays, on human samples?
   • Unfortunately at this time we have not tested, or received customer feedback on our MMP8 antibodies for use in blocking or neutralizing assays. If you were planning on testing any of our MMP8 antibodies for neutralizing or blocking capabilities, we would recommend our Innovator’s Reward Program. Under the terms of this program we would be happy to provide a credit for a free vial in exchange for new data on this previously untested or reported application. Please submit this data in the form of an online review. Additional Innovators Reward Program information can be found using this link.
2. We would like to stain paraffin embedded human intestinal tissue for Calprotectin. One question will be which cells express the majority of Calprotectin - invading neutrophils vs epithelium for example. The antibody list you provide is quite extensive. Which antibody would you recommend for our purpose? Which tissue would you recommend as positive control.
   • Nine of our antibodies to Calprotectin have been validated for IHC-P with human tissue, and you can see all of these products. A number of our IHC images were generated using human spleen samples, and as such this may be a good choice of a positive control tissue. We do sell a human spleen slide product, which is suitable for IHC-P. Unfortunately I do not have an in-depth knowledge of Calprotectin, however the image shown for our antibody with catalogue number NBP1-02826 demonstrates clear staining of neutrophils. The following paper also suggests that the protein is abundant in neutrophils: PMID 11435495.

Bioinformatics

• Gene Symbol: S100A8
• Uniprot:
  • P27005()