Human/Mouse c-Myc Antibody

Detection of Human c-Myc by Western Blot. Western blot shows lysates of LNCaP human prostate cancer cell line and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for c-Myc at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse c‑Myc by Western Blot. Western blot shows lysates of BaF3 mouse pro-B cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for c-Myc at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of c‑Myc-regulated Genes by Chromatin Immunoprecipitation. HeLa human cervical epithelial carcinoma cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. c-Myc/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thep21promoter was detected by standard PCR.

c‑Myc in D3 Mouse Stem Cells. c-Myc was detected in immersion fixed D3 mouse embryonic stem cell line using Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

c‑Myc in BG01V Human Stem Cells. c-Myc was detected in immersion fixed BG01V human embryonic stem cells using 10 µg/mL Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human c‑Myc by Simple Western^TM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for c-Myc at approximately 78 kDa (as indicated) using 20 µg/mL of Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Shows Human c‑Myc Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and c-Myc knockout HEK293T cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for c-Myc at approximately 52 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse c-Myc by Western Blot DON significantly inhibits xenograft tumor growth.(A) SK-N-AS tumors were treated with DON at 100 mg/kg or water by i.p. twice weekly. Weight loss in DON-treated mice reduced the treatment cohort to 2 mice indicated by (2) at later timepoints. (B) SK-N-FI tumors were treated with DON at 100 mg/kg, 50 mg/kg or water by i.p. twice weekly or 100mg/kg once a week (100mg/kg; 1x/wk). The * indicates p< 0.05 between control and 50 mg/kg, while ** indicates p<0.05 between control and 100mg/kg; 1x/wk, Student’s t test. (C and D) For SK-N-BE2 and IMR32 subcutaneous xenografts, mice were treated with DON at 50 mg/kg or water control twice weekly by i.p. injections. The indicated tumor cell line was grown to 200 mm3 prior to initiation of treatment and tumor size is given as a percent relative to original tumor volume (% RTV). (E) Western blot analysis of N-Myc and c-Myc expression in a panel of NBL cell lines with beta -tubulin as a loading control. (F) A composite graph of SK-N-FI, SK-N-BE2 and IMR32 tumors treated with DON 50mg/kg or water control twice weekly. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25615615), licensed under a CC-BY license. Not internally tested by R&D Systems.