Detection of Human 53BP1 by Western Blot.
Western blot shows lysates of HEK293T human embryonic kidney cell line and ZR‑75 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1877) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for 53BP1 at approximately 350 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
53BP1 in Rat Kidney.
53BP1 was detected in perfusion fixed frozen sections of rat kidney using Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1877) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei of epithelial cells in convoluted tubules. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Western Blot Shows Human 53BP1 Specificity by Using Knockout Cell Line.
Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and 53BP1 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1877) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for 53BP1 at approximately 350 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.
The p53-binding protein 1 (53BP1) localizes to sites of damaged DNA and is phosphorylated by the ATM checkpoint kinase. 53BP1 is required for phosphorylation of other downstream ATM substrates, such as p53 and SMC-1, and therefore aids in nucleating a DNA damage response protein complex. Over aa 1614-1972, human 53BP1 shares 97% aa sequence identity with rat and mouse 53BP1.
p53 Binding Protein 1
Entrez Gene IDs:
7158 (Human); 27223 (Mouse); 296099 (Rat)
53BP1; MGC138366; p202; p53-binding protein 1; p53BP1,53BP1FLJ41424; TP53BP1; tumor protein 53-binding protein, 1; tumor protein p53 binding protein 1; tumor protein p53-binding protein, 1; tumor suppressor p53-binding protein 1
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