Purified anti-53BP1 Antibody

Pricing & Availability
Clone
W17184B (See other available formats)
Regulatory Status
RUO
Other Names
Tumor Protein P53 Binding Protein 1, TP53-Binding Protein 1, P53-Binding Protein 1, P53BP1, Tumor Suppressor P53-Binding Protein 1, TDRD30, P202
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
W17184B_PURE_53BP1_Antibody_1_101719.png
Whole cell extracts (15 µg protein) from the indicated cell lines were resolved on a 3-8% Tris-Acetate gel, transferred to PVDF and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-53BP1 Antibody (clone W17184B) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG Antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker. Lysates were loaded in order of descending TP53BP1 expression (Source: Human Protein Atals).
  • W17184B_PURE_53BP1_Antibody_1_101719.png
    Whole cell extracts (15 µg protein) from the indicated cell lines were resolved on a 3-8% Tris-Acetate gel, transferred to PVDF and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-53BP1 Antibody (clone W17184B) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG Antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker. Lysates were loaded in order of descending TP53BP1 expression (Source: Human Protein Atals).
  • W17184B_PURE_53BP1_Antibody_2_101719.png
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with a 1.0 µg/mL (1:500 dilution) of either Purified Rat IgG2a, κ Isotype Ctrl Antibody (Cat. No. 400502, panel A) or Purified anti-53BP1 Antibody (clone W17184B, panel B) overnight at 4°C, followed by incubation with Alexa Fluor® 594 Goat anti-rat IgG Antibody (Cat. No. 405422) at 2.0 µg/mL. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
Cat # Size Price Quantity Check Availability Save
933001 25 µg 101 CHF
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933002 100 µg 253 CHF
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Description

p53-binding protein 1 (53BP1) was first discovered as an interacting partner of p53 tumor suppressor protein in a yeast two-hybrid screening. 53BP1 binds to p53 through a BRCA1 C-terminal (BRCT) domain and enhances p53-dependent transcriptional activation. 53BP1 has been studied extensively for its function in the regulation of DNA double-strand break (DSB) repair pathway. Additionally, it promotes non-homologous end joining (NHEJ) while inhibiting homologous recombination (HR) by preventing DNA end resection. Upon DSB, the ubiquitin ligase RNF168 ubiquitylates lysine 15 on histone H2A/H2AX, which serves as a binding site for 53BP1. 53BP1 is then phosphorylated by ATM, which facilitates the recruitment of downstream effector proteins, including PTIP and RIF1. 53BP1-mediated NHEJ is essential for V(D)J recombination and class switch recombination.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial recombinant human 53BP1 protein.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified

Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone recognizes both human and mouse 53BP1.

This clone was tested for ICC using both Triton X-100 and methanol permeabilization with both methods showing correct staining.

This clone failed IP testing.

Predicted MW of 53BP1 is 214 kD but the observed MW is ~350kD. An identical observed MW was noted when comparing to other control antibodies on the market. Similar differences in predicted and observed molecular weights are reported with other antibodies on the market.

Product Citations
  1. Zhang L, et al. 2022. Nat Commun. 13:360. PubMed
RRID
AB_2820202 (BioLegend Cat. No. 933001)
AB_2820202 (BioLegend Cat. No. 933002)

Antigen Details

Structure
53BP1 is a 1972 amino acid protein with a predicted molecular weight of 214 kD.
Distribution

Nucleus/Ubiquitously expressed

Function
DNA damage response
Biology Area
DNA Repair/Replication
Antigen References
  1. Iwabuchi K, et al. 1994. Proc Natl Acad Sci USA. 91:6098.
  2. Wang B, et al. 2002. Science. 298:1435.
  3. Difilippantonio S, et al. 2008. Nature. 456:529.
  4. Bunting SF, et al. 2010. Cell. 141:243.
  5. Cuella-Martin R, et al. 2016. Molecular Cell. 64:51.
Gene ID
7158 View all products for this Gene ID
UniProt
View information about 53BP1 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 10.17.2019

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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