| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP2-74647 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP3-43938 | |||
| BSA Free | NBP3-07990-25ul | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP2-93270-0.02ml | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF2747-SP | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | SCLU00 | |||
| Carrier Free | PDCLU00 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | MAB2937-SP | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF2937-SP | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF7084-SP | |||
![Western Blot: TRPM4 Antibody (OTI10H5) [NBP1-48036] - Analysis of extracts (35ug) from 9 different cell lines by using anti-TRPM4 monoclonal antibody.](http://images.novusbio.com/fullsize/TRPM4-Antibody-10H5-Western-Blot-NBP1-48036-img0007.jpg "Western Blot: TRPM4 Antibody (OTI10H5) [NBP1-48036] - Analysis of extracts (35ug) from 9 different cell lines by using anti-TRPM4 monoclonal antibody.")
![Immunohistochemistry-Paraffin: TRPM4 Antibody (OTI10H5) [NBP1-48036] - Human endometrial carcinoma tissue section. Fixed paraffin embedded sections were deparaffinized. EDTA was used for antigen retrieval. Samples were incubated O/N in 4 C with the primary antibody (1:200 dilution). IHC-P image submitted by a verified review.](http://images.novusbio.com/fullsize/TRPM4-Antibody-OTI10H5-Immunohistochemistry-Paraffin-NBP1-48036-img0009.jpg)
![Western Blot: TRPM4 Antibody (10H5) [NBP1-48036] - HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TRPM4 (Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TRPM4.](http://images.novusbio.com/fullsize/TRPM4-Antibody-10H5-Western-Blot-NBP1-48036-img0001.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (10H5) [NBP1-48036] - Staining of paraffin-embedded Carcinoma of Human liver tissue using anti-TRPM4 mouse monoclonal antibody.](http://images.novusbio.com/fullsize/TRPM4-Antibody-10H5-Immunohistochemistry-Paraffin-NBP1-48036-img0002.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (10H5) [NBP1-48036] - Staining of paraffin-embedded Human Kidney tissue using anti-TRPM4 mouse monoclonal antibody.](http://images.novusbio.com/fullsize/TRPM4-Antibody-10H5-Immunohistochemistry-Paraffin-NBP1-48036-img0003.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (10H5) [NBP1-48036] - Staining of paraffin-embedded Human liver tissue using anti-TRPM4 mouse monoclonal antibody.](http://images.novusbio.com/fullsize/TRPM4-Antibody-10H5-Immunohistochemistry-Paraffin-NBP1-48036-img0004.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (10H5) [NBP1-48036] - Staining of paraffin-embedded Human Ovary tissue using anti-TRPM4 mouse monoclonal antibody.](http://images.novusbio.com/fullsize/TRPM4-Antibody-10H5-Immunohistochemistry-Paraffin-NBP1-48036-img0005.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (10H5) [NBP1-48036] - Staining of paraffin-embedded Human pancreas tissue using anti-TRPM4 mouse monoclonal antibody.](http://images.novusbio.com/fullsize/TRPM4-Antibody-10H5-Immunohistochemistry-Paraffin-NBP1-48036-img0006.jpg)
![Immunohistochemistry: TRPM4 Antibody (OTI10H5) [NBP1-48036] - Human heart tissue was blocked with 10% serum and stained with the primary antibody at 1:100 dilution overnight at 4 degrees. A secondary FITC-conjugated anti-mouse antibody was used in 1:400 dilution for 2 hours at room temperature. IHC image submitted by a verified customer review.](http://images.novusbio.com/fullsize/TRPM4-Antibody-OTI10H5-Immunohistochemistry-NBP1-48036-img0008.jpg)
![TRPM4 Antibody (OTI10H5)[NBP1-48036]Simple Western� analysis of endogenous protein ACTN1 from HELA lysates (0.2 mg/mL) using ACTN1 Mouse Monoclonal Antibody . The virtual lane view (left) shows the target (as indicated) at 1:50 dilution of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at a 1:50 dilution of primary antibody. This experiment was performed under reducing conditions on the Jess� Simple Western instrument from ProteinSimple, a Bio-Techne brand, using the 12�230 kDa Separation Module.](http://images.novusbio.com/fullsize/nbp1-48036_mouse-trpm4-mab-oti10h5-simple-western-1311202512362210.jpg%253fstore=america&image-type=image)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (CL11214) [NBP3-07990] - Analysis in human duodenum and pancreas tissues using NBP3-07990 antibody. Corresponding TRPM4 RNA-seq data are presented for the same tissues.](http://images.novusbio.com/fullsize/TRPM4-Antibody-CL11214-Immunohistochemistry-Paraffin-NBP3-07990-img0001.jpg "<b>Orthogonal Strategies Validation. </b>Immunohistochemistry-Paraffin: TRPM4 Antibody (CL11214) [NBP3-07990] - Analysis in human duodenum and pancreas tissues using NBP3-07990 antibody. Corresponding TRPM4 RNA-seq data are presented for the same tissues.")
![<b>Orthogonal Strategies Validation. </b>Western Blot: TRPM4 Antibody (CL11214) [NBP3-07990] - Analysis in human cell line RT-4.](http://images.novusbio.com/fullsize/TRPM4-Antibody-CL11214-Western-Blot-NBP3-07990-img0007.jpg)
![Immunocytochemistry/Immunofluorescence: TRPM4 Antibody (CL11214) [NBP3-07990] - Staining of MCF7 cells using the Anti-TRPM4 monoclonal antibody, showing specific staining in the plasma membrane in green. Microtubule- and nuclear probes are visualized in red and blue, respectively (where available).](http://images.novusbio.com/fullsize/TRPM4-Antibody-CL11214-Immunocytochemistry-Immunofluorescence-NBP3-07990-img0006.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (CL11214) [NBP3-07990] - Staining of human prostate shows moderate membranous positivity in glandular cells.](http://images.novusbio.com/fullsize/TRPM4-Antibody-CL11214-Immunohistochemistry-Paraffin-NBP3-07990-img0005.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (CL11214) [NBP3-07990] - Staining of human duodenum shows strong membranous positivity in glandular cells.](http://images.novusbio.com/fullsize/TRPM4-Antibody-CL11214-Immunohistochemistry-Paraffin-NBP3-07990-img0002.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (CL11214) [NBP3-07990] - Staining of human kidney shows moderate to strong membranous positivity in cells in tubules.](http://images.novusbio.com/fullsize/TRPM4-Antibody-CL11214-Immunohistochemistry-Paraffin-NBP3-07990-img0003.jpg)
![Immunohistochemistry-Paraffin: TRPM4 Antibody (CL11214) [NBP3-07990] - Staining of human pancreas shows no positivity in exocrine glandular cells as expected.](http://images.novusbio.com/fullsize/TRPM4-Antibody-CL11214-Immunohistochemistry-Paraffin-NBP3-07990-img0004.jpg)
![Western Blot: TRPM4 Antibody [NBP2-93270] - Western blot analysis of extracts of Rat brain, using TRPM4 antibody (NBP2-93270) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 60s.](http://images.novusbio.com/fullsize/TRPM4 Antibody-Western Blot-NBP2-93270-img0001.jpg "Western Blot: TRPM4 Antibody [NBP2-93270] - Western blot analysis of extracts of Rat brain, using TRPM4 antibody (NBP2-93270) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 60s.")
![Conservation of clusterin‐dependent alpha ‐synuclein preformed fibrils (pffs) uptake from mouse to human astrocytes. (a) Representative image of hiAstrocytes (27 days of differentiation) treated with * alpha ‐synuclein pffs for 48 hr. Scale bar 10 μm. (b) Cell lysates from hiAstrocytes (27 days of differentiation) treated with alpha ‐synuclein pffs, or monomeric alpha ‐synuclein (m), for 48 hr were subjected to immunoblotting using alpha ‐synuclein and GAPDH antibodies. Asterisks in the alpha ‐synuclein immunoblot indicate alpha ‐synuclein nonspecific bands. (c) Medium from hiAstrocytes (27 days of differentiation) treated with alpha ‐synuclein pffs, or monomeric alpha ‐synuclein (M), for 48 hr were subjected to immunoblotting using clusterin antibody. Ponceau‐S staining was used as loading control. (d) Quantification of clusterin is normalized to GAPDH protein of cell lysates. Data are representative of three independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0446. (e) Cell lysates from hiAstrocytes (31 days of differentiation) transfected with negative control (NC) or clusterin siRNA for 24 hr were subjected to immunoblotting using clusterin and GAPDH antibodies. Asterisk in the clusterin immunoblot indicates clusterin nonspecific band. (f) Quantification of clusterin precursor is normalized to GAPDH. Data are representative of three independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0165. (g) Representative images of hiAstrocytes (35 days of differentiation) transfected with NC or clusterin siRNA and treated with * alpha ‐synuclein pffs for 24 hr. Scale bar 10 μm. (h) Quantification of * alpha ‐synuclein pffs is shown as mean of fluorescence intensity from three independent experiments (~50 cells per experiment). Quantification of * alpha ‐synuclein pffs is calculated as fluorescence intensity divided by the cell area (μm2). Data are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0178. Individual points of the graphs represent each single experiment [Color figure can be viewed at wileyonlinelibrary.com] Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af2747_mouse-clusterin-affinity-purified-polyclonal-ab-41202412402536.jpg)
![<b>Genetic Strategies Validation. </b>Extracellular clusterin limits the uptake of alpha ‐synuclein preformed fibrils (pffs) by murine primary astrocytes. (a) Cell lysates from clusterin wild‐type (WT) and knock‐out (KO) astrocytes were subjected to immunoblotting using clusterin and GAPDH antibodies. Asterisk indicates clusterin precursor. (b) Maximum intensity Z‐projection confocal images of clusterin WT and KO astrocytes treated with * alpha ‐synuclein pffs for 4 hr. Scale bar 5 μm. (c) Quantification of * alpha ‐synuclein pffs is shown as mean of fluorescence intensity from six independent experiments (~50 cells per experiment). Quantification of * alpha ‐synuclein pffs is calculated as fluorescence intensity divided by the cell area (μm2). Data are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0315. (d) Cell lysates from clusterin WT and KO primary astrocytes treated with pffs for 16 hr were subjected to immunoblotting using alpha ‐synuclein and GAPDH antibodies. (e) Quantification of intracellular alpha ‐synuclein is normalized to GAPDH protein. Data are representative of eight independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0239. (f) Cell lysates from clusterin KO primary astrocytes treated with pffs for 16 hr in a medium containing or not containing extracellular clusterin were subjected to immunoblotting using alpha ‐synuclein, clusterin and GAPDH antibodies. (g) Quantification of intracellular alpha ‐synuclein is normalized to GAPDH protein. Data are representative of four independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0134. (h) Cell lysates from clusterin WT and KO primary astrocytes treated with pffs and chloroquine (CQ) for 16 hr were subjected to immunoblotting using alpha ‐synuclein and GAPDH antibodies. (i) Quantification of intracellular alpha ‐synuclein normalized to GAPDH protein. Data are representative of six independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0377. Individual points of the graphs represent each single experiment [Color figure can be viewed at wileyonlinelibrary.com] Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af2747_mouse-clusterin-affinity-purified-polyclonal-ab-41202412403576.jpg)
![Clusterin protein levels are increased in two different animal models of Parkinson's disease. (a) Lysates from contralateral and ipsilateral ventral midbrain of mice injected with adeno‐associated virus (AAV)‐h alpha ‐synuclein and sacrificed after 8 weeks were subjected to immunoblotting using clusterin and GAPDH antibodies. (b) Quantification of total cleaved clusterin is normalized to GAPDH protein. Data are representative of five animals and are expressed as the mean ± SD. Data were analyzed using one‐sample t test; *p = .0253. (c) Representative images of clusterin‐GFAP double labeling in the ipsilateral and contralateral striatum of mice injected with AAV‐h alpha ‐synuclein and sacrificed after 8 weeks. Scale bar 20 μm. (d) Representative images of clusterin‐S100 beta double labeling in the substantia nigra of mice injected with alpha ‐synuclein preformed fibrils (pffs) and sacrificed after 4 weeks. Scale bar 20 μm. (e) Representative images of clusterin‐S100 beta double labeling in the striatum of mice injected with alpha ‐synuclein pffs and sacrificed after 4 weeks. Scale bar 20 μm [Color figure can be viewed at wileyonlinelibrary.com] Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af2747_mouse-clusterin-affinity-purified-polyclonal-ab-41202412403564.jpg)
![Conservation of clusterin‐dependent alpha ‐synuclein preformed fibrils (pffs) uptake from mouse to human astrocytes. (a) Representative image of hiAstrocytes (27 days of differentiation) treated with * alpha ‐synuclein pffs for 48 hr. Scale bar 10 μm. (b) Cell lysates from hiAstrocytes (27 days of differentiation) treated with alpha ‐synuclein pffs, or monomeric alpha ‐synuclein (m), for 48 hr were subjected to immunoblotting using alpha ‐synuclein and GAPDH antibodies. Asterisks in the alpha ‐synuclein immunoblot indicate alpha ‐synuclein nonspecific bands. (c) Medium from hiAstrocytes (27 days of differentiation) treated with alpha ‐synuclein pffs, or monomeric alpha ‐synuclein (M), for 48 hr were subjected to immunoblotting using clusterin antibody. Ponceau‐S staining was used as loading control. (d) Quantification of clusterin is normalized to GAPDH protein of cell lysates. Data are representative of three independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0446. (e) Cell lysates from hiAstrocytes (31 days of differentiation) transfected with negative control (NC) or clusterin siRNA for 24 hr were subjected to immunoblotting using clusterin and GAPDH antibodies. Asterisk in the clusterin immunoblot indicates clusterin nonspecific band. (f) Quantification of clusterin precursor is normalized to GAPDH. Data are representative of three independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0165. (g) Representative images of hiAstrocytes (35 days of differentiation) transfected with NC or clusterin siRNA and treated with * alpha ‐synuclein pffs for 24 hr. Scale bar 10 μm. (h) Quantification of * alpha ‐synuclein pffs is shown as mean of fluorescence intensity from three independent experiments (~50 cells per experiment). Quantification of * alpha ‐synuclein pffs is calculated as fluorescence intensity divided by the cell area (μm2). Data are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0178. Individual points of the graphs represent each single experiment [Color figure can be viewed at wileyonlinelibrary.com] Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af2747_mouse-clusterin-affinity-purified-polyclonal-ab-4120241240247.jpg)
![<b>Genetic Strategies Validation. </b>Extracellular clusterin limits the uptake of alpha ‐synuclein preformed fibrils (pffs) by murine primary astrocytes. (a) Cell lysates from clusterin wild‐type (WT) and knock‐out (KO) astrocytes were subjected to immunoblotting using clusterin and GAPDH antibodies. Asterisk indicates clusterin precursor. (b) Maximum intensity Z‐projection confocal images of clusterin WT and KO astrocytes treated with * alpha ‐synuclein pffs for 4 hr. Scale bar 5 μm. (c) Quantification of * alpha ‐synuclein pffs is shown as mean of fluorescence intensity from six independent experiments (~50 cells per experiment). Quantification of * alpha ‐synuclein pffs is calculated as fluorescence intensity divided by the cell area (μm2). Data are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0315. (d) Cell lysates from clusterin WT and KO primary astrocytes treated with pffs for 16 hr were subjected to immunoblotting using alpha ‐synuclein and GAPDH antibodies. (e) Quantification of intracellular alpha ‐synuclein is normalized to GAPDH protein. Data are representative of eight independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0239. (f) Cell lysates from clusterin KO primary astrocytes treated with pffs for 16 hr in a medium containing or not containing extracellular clusterin were subjected to immunoblotting using alpha ‐synuclein, clusterin and GAPDH antibodies. (g) Quantification of intracellular alpha ‐synuclein is normalized to GAPDH protein. Data are representative of four independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0134. (h) Cell lysates from clusterin WT and KO primary astrocytes treated with pffs and chloroquine (CQ) for 16 hr were subjected to immunoblotting using alpha ‐synuclein and GAPDH antibodies. (i) Quantification of intracellular alpha ‐synuclein normalized to GAPDH protein. Data are representative of six independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; *p = .0377. Individual points of the graphs represent each single experiment [Color figure can be viewed at wileyonlinelibrary.com] Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af2747_mouse-clusterin-affinity-purified-polyclonal-ab-41202412402523.jpg)
![N/A Clusterin/APOJ [HRP]](http://images.novusbio.com/fullsize2/DATA_Clusterin_DCLU00_ELISA_631.jpg "N/A Clusterin/APOJ [HRP]")
![N/A Clusterin/APOJ [HRP]](http://images.novusbio.com/fullsize2/Clusterin_DCLU00_ELISA_65.jpg)
![Extracellular clusterin interacts with alpha ‐synuclein preformed fibrils (pffs). (a) Medium from primary astrocytes treated with alpha ‐synuclein pffs for 16 hr, or with monomeric alpha ‐synuclein (m), were subjected to immunoblotting using clusterin antibody. Ponceau‐S staining was used as a loading control. (b) Quantification of clusterin is normalized to GAPDH protein of cell lysates. Data are from eight independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; **p = .0028. (c) alpha ‐Synuclein pffs isolated by centrifugation from total medium of primary astrocytes treated with alpha ‐synuclein pffs, or monomeric alpha ‐synuclein (M), for 16 hr were subjected to immunoblotting using clusterin and alpha ‐synuclein antibodies. Asterisk in the alpha ‐synuclein immunoblot indicates clusterin bands still present after Clusterin antibody stripping. (d) Clusterin immunoprecipitated from the medium of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr was subjected to immunoblotting using an alpha ‐synuclein antibody. (e) Quantification of clusterin— alpha ‐synuclein pffs interaction has been obtained by normalization of alpha ‐synuclein pffs bound to clusterin for alpha ‐synuclein pffs bound to protein‐G beads. Data are representative of three independent experiments and are expressed as the mean ± SD. Data were analyzed using one‐sample t test; *p = .0493. (f) Z‐stack frames of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr and stained for alpha ‐synuclein (purple), clusterin (green), cadherin (red), and nuclei with DAPI (blue). Scale bar 10 μm. (g) Maximum intensity Z‐projection confocal images of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr and stained for alpha ‐synuclein (purple), clusterin (red), Early Endosome Antigen 1 (EEA1) (green) and nuclei with DAPI (blue). Scale bar 10 μm. Individual points of the graphs represent each single experiment [Color figure can be viewed at wileyonlinelibrary.com] Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af2937_human-clusterin-affinity-purified-polyclonal-ab-41202412401226.jpg)
![Extracellular clusterin interacts with alpha ‐synuclein preformed fibrils (pffs). (a) Medium from primary astrocytes treated with alpha ‐synuclein pffs for 16 hr, or with monomeric alpha ‐synuclein (m), were subjected to immunoblotting using clusterin antibody. Ponceau‐S staining was used as a loading control. (b) Quantification of clusterin is normalized to GAPDH protein of cell lysates. Data are from eight independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; **p = .0028. (c) alpha ‐Synuclein pffs isolated by centrifugation from total medium of primary astrocytes treated with alpha ‐synuclein pffs, or monomeric alpha ‐synuclein (M), for 16 hr were subjected to immunoblotting using clusterin and alpha ‐synuclein antibodies. Asterisk in the alpha ‐synuclein immunoblot indicates clusterin bands still present after Clusterin antibody stripping. (d) Clusterin immunoprecipitated from the medium of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr was subjected to immunoblotting using an alpha ‐synuclein antibody. (e) Quantification of clusterin— alpha ‐synuclein pffs interaction has been obtained by normalization of alpha ‐synuclein pffs bound to clusterin for alpha ‐synuclein pffs bound to protein‐G beads. Data are representative of three independent experiments and are expressed as the mean ± SD. Data were analyzed using one‐sample t test; *p = .0493. (f) Z‐stack frames of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr and stained for alpha ‐synuclein (purple), clusterin (green), cadherin (red), and nuclei with DAPI (blue). Scale bar 10 μm. (g) Maximum intensity Z‐projection confocal images of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr and stained for alpha ‐synuclein (purple), clusterin (red), Early Endosome Antigen 1 (EEA1) (green) and nuclei with DAPI (blue). Scale bar 10 μm. Individual points of the graphs represent each single experiment [Color figure can be viewed at wileyonlinelibrary.com] Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af2937_human-clusterin-affinity-purified-polyclonal-ab-41202412401245.jpg)
![Extracellular clusterin interacts with alpha ‐synuclein preformed fibrils (pffs). (a) Medium from primary astrocytes treated with alpha ‐synuclein pffs for 16 hr, or with monomeric alpha ‐synuclein (m), were subjected to immunoblotting using clusterin antibody. Ponceau‐S staining was used as a loading control. (b) Quantification of clusterin is normalized to GAPDH protein of cell lysates. Data are from eight independent experiments and are expressed as the mean ± SD. Data were analyzed using unpaired t test; **p = .0028. (c) alpha ‐Synuclein pffs isolated by centrifugation from total medium of primary astrocytes treated with alpha ‐synuclein pffs, or monomeric alpha ‐synuclein (M), for 16 hr were subjected to immunoblotting using clusterin and alpha ‐synuclein antibodies. Asterisk in the alpha ‐synuclein immunoblot indicates clusterin bands still present after Clusterin antibody stripping. (d) Clusterin immunoprecipitated from the medium of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr was subjected to immunoblotting using an alpha ‐synuclein antibody. (e) Quantification of clusterin— alpha ‐synuclein pffs interaction has been obtained by normalization of alpha ‐synuclein pffs bound to clusterin for alpha ‐synuclein pffs bound to protein‐G beads. Data are representative of three independent experiments and are expressed as the mean ± SD. Data were analyzed using one‐sample t test; *p = .0493. (f) Z‐stack frames of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr and stained for alpha ‐synuclein (purple), clusterin (green), cadherin (red), and nuclei with DAPI (blue). Scale bar 10 μm. (g) Maximum intensity Z‐projection confocal images of primary astrocytes treated with alpha ‐synuclein pffs for 16 hr and stained for alpha ‐synuclein (purple), clusterin (red), Early Endosome Antigen 1 (EEA1) (green) and nuclei with DAPI (blue). Scale bar 10 μm. Individual points of the graphs represent each single experiment [Color figure can be viewed at wileyonlinelibrary.com] Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af2937_human-clusterin-affinity-purified-polyclonal-ab-41202412402530.jpg)
![N/A Clusterin/APOJ [HRP]](http://images.novusbio.com/fullsize2/DATA_Clusterin_MCLU00_ELISA_957.jpg "N/A Clusterin/APOJ [HRP]")
![N/A Clusterin/APOJ [HRP]](http://images.novusbio.com/fullsize2/Clusterin_MCLU00_ELISA_253.jpg)
![N/A Clusterin/APOJ [Biotin]](http://images.novusbio.com/fullsize2/DATA_Clusterin_DY5874_ELISA_2139.jpg "N/A Clusterin/APOJ [Biotin]")
