Human Clusterin Isoform 1 Antibody

Detection of Human Clusterin by Western Blot. Western blot shows lysates of human liver tissue and human serum. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Clusterin Isoform 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2937) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Clusterin Precusor at approximately 60-65 kDa and Clusterin a and beta chains at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Clusterin in Human Alzheimer's Disease Brain. Clusterin was detected in immersion fixed paraffin-embedded of human Alzheimer's disease brain using Human Clusterin Isoform 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2937) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Clusterin/APOJ by Western Blot 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 (https://pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Clusterin/APOJ by Western Blot 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 (https://pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Clusterin/APOJ by Western Blot 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 (https://pubmed.ncbi.nlm.nih.gov/33045109), licensed under a CC-BY license. Not internally tested by R&D Systems.