Human 53BP1 Antibody

R&D Systems | Catalog # AF1877

R&D Systems

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, Immunohistochemistry, Western Blot, Simple Western

Cited:

Immunohistochemistry, Immunocytochemistry, FISH

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all 53BP1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human 53BP1
Ala1614-His1972
Accession # Q12888

Specificity

Detects human 53BP1 in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human 53BP1 Antibody

Detection of Human 53BP1 antibody by Western Blot.

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 antibody in Rat Kidney by Immunohistochemistry (IHC-Fr).

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.

Detection of Human 53BP1 by Simple WesternTM.

Left: Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 1 mg/ml. A specific band was detected for 53BP1 at approximately 352 kDa (as indicated) using both 2 µg/ml and 10 µg/ml of Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # af1877) followed by HRP-conjugated Donkey Anti-Goat Secondary Antibody (Catalog # 043-522-2). This experiment was conducted under reducing conditions and using the 66-440kDa separation system. Right: Simple Western electropherogram showing the same Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # af1877) tested at 2 µg/ml (blue line) and 10 µg/ml (green line) in the HepG2 human hepatocellular carcinoma cell line.
Western Blot Shows Human 53BP1 Antibody Specificity by Using Knockout Cell Line.

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.
Detection of 53BP1 by Immunocytochemistry/ Immunofluorescence

Detection of 53BP1 by Immunocytochemistry/ Immunofluorescence

Visualising RAD51 accumulations. (A) U2OS cells were treated with non-target control siRNA (NTC) or siRNA targeting USP48 (USP48 depleted) for 72 h. Cells were treated with EdU (blue) prior to irradiation (2 Gy) and were then allowed to recover for 1 h prior to fixation. Cells were immunostained for RAD51 (green), BRCA1 (white) and 53BP1 (magenta) then prepared using the ExM method. Post-expansion images of late S-phase nuclei are shown. Boxes indicate regions shown in the magnified images on the right. Scale bars: 10 μm (main images) and 2 μm (selected regions), equivalent to ∼2.5 μm and 500 nm pre-expansion, respectively. (B) U2OS cells were treated with non-target control siRNA or siRNA targeting USP48 for 72 h. Cells were treated with EdU (blue) prior to irradiation (2 Gy) and were then allowed to recover for 1 h prior to fixation. Cells were immunostained for RAD51 (green) and 53BP1 (magenta), then prepared using the ExM method. Post-expansion images of late S-phase nuclei are shown. Boxes indicate regions shown in the magnified images on the right. Scale bars: 10 μm (main images) and 2 μm (selected regions), equivalent to ∼2.5 μm and 500 nm pre-expansion, respectively. (C) Examples of co-enriched structures with RAD51 (green) and 53BP1 (magenta) from late S-phase nuclei are shown. Scale bars: 2 μm (equivalent to ∼500 nm pre-expansion). (D) Examples of structures co-enriched with BRCA1 (yellow), RAD51 (green) and 53BP1 (magenta) from late S-phase nuclei are shown. Scale bars: 2 μm (equivalent to ∼500 nm pre-expansion). Images in A and C are representative of three experiments. Images in B and D are representative of two experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35748225), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human 53BP1 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of rat kidney

Knockout Validated

0.5 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and HeLa with knockout of p53BP1

Simple Western

2-10 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line

Western Blot

1 µg/mL
Sample: HEK293T human embryonic kidney cell line and ZR‑75 human breast cancer cell line

Reviewed Applications

Read 1 review rated 5 using AF1877 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: 53BP1

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. 

Long Name

p53 Binding Protein 1

Alternate Names

p202, TP53BP1

Entrez Gene IDs

7158 (Human); 27223 (Mouse); 296099 (Rat)

Gene Symbol

TP53BP1

UniProt

Q12888

Additional 53BP1 Products

Product Documents for Human 53BP1 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human 53BP1 Antibody

For research use only

Related Research Areas

p53 Pathway

Citations for Human 53BP1 Antibody

Customer Reviews for Human 53BP1 Antibody (1)

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  • Human 53BP1 Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: U2OS
    Species: Human
    Verified Customer | Posted 12/30/2019
    4% paraformaldehyde fixed U2OS cells were stained with anti-53BP1 followed by Alexa488 anti-goat secondary antibody.
    Human 53BP1 Antibody AF1877

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Protocols

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