Human/Mouse WDR5 Antibody

Detection of Human WDR5 by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line. Gels were loaded with 30 µg of whole cell lysate (WCL), 20 µg of cytoplasmic (Cyto), and 10 µg of nuclear extracts (Nuc). PVDF membrane was probed with 1 µg/mL Goat Anti-Human/Mouse WDR5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5810) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band for WDR5 was detected at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

WDR5 in Human B-Cell Lymphoma.

WDR5 was detected in immersion fixed paraffin-embedded sections of human B-cell lymphoma using Goat Anti-Human/Mouse WDR5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5810) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human WDR5 by Immunocytochemistry/Immunofluorescence

SRY cooperates with WDR5 to induce Sox9 expression.(A) Immunofluorescence analysis of HA-tagged SRY and WDR5 in LNCaP cells. (B) ChIP-reChIP (anti-HA antibody ChIP followed by anti-WDR5 antibody ChIP) analysis of HA-tagged SRY and WDR5 on Sox9 promoter, or Sox9 promoter proximal upstream region. Graphs show mean ± SD, n = 3. (C) Coimmunoprecipitation of WDR5 and HA-tagged SRY from LNCaP cells. (D) Quantitative real time PCR analysis of Sox9 levels from LNCaP cells containing SF1, SF1+WDR5, SF1+SRY, or SF1+SRY+WDR5. Graphs show mean ± SD, n = 3. *P<0.05, **P<0.01 compared to SF1 control; Student's t-test. (E) Quantitative real time PCR analysis of beta-catenin levels as in (D). Graphs show mean ± SD, n = 3. *P<0.05, **P<0.01 compared to SF1 control; Student's t-test. (F) Western blot analysis of cellular lysate from cells as in (D). (G) Western blot analyses of cellular extracts from WDR5i-1 and WDR5i-2 or scrambled control (Ctrl) LNCaP cells with indicated antibodies. (H) WDR5 gene expression analysis by Q-RT-PCR of RNA from WDR5i-1, WDR5i-2 and scrambled control (Ctrl) LNCaP cells. Graphs show mean ± SD, n = 3. *P<0.05 compared to the scrambled control, Student's t-test. (I) Sox9 gene expression analysis by Q-RT-PCR of RNA from WDR5i-1, WDR5i-2 and scrambled control (Ctrl) LNCaP cells. Graphs show mean±SD, n = 3. **P<0.01 compared to the scrambled control, Student's t-test. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0034327), licensed under a CC-BY license. Not internally tested by R&D Systems.