Overview

  • Product name

  • Description

    Rabbit polyclonal to 53BP1
  • Host species

    Rabbit
  • Specificity

    Recognises 53BP1
  • Tested applications

    Suitable for: In situ hybridization, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human 53BP1 (C terminal). Conjugated to a carrier protein.
    Database link: Q12888

  • Positive control

    • WB: Neuro2A, C8D30, NIH-3T3, 293T, A431, HeLa, HepG2 whole cell extracts. IHC-P: Human colon cancer tissue, human brain tissue, human breast carcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab21083 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
In situ hybridization Use at an assay dependent concentration. PubMed: 23370393
WB 1/500 - 1/3000. Detects a band of approximately 350 kDa (predicted molecular weight: 220 kDa).
IHC-P 1/1000 - 1/4000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    May have a role in checkpoint signaling during mitosis. Enhances TP53-mediated transcriptional activation. Plays a role in the response to DNA damage.
  • Involvement in disease

    Note=A chromosomal aberration involving TP53BP1 is found in a form of myeloproliferative disorder chronic with eosinophilia. Translocation t(5;15)(q33;q22) with PDGFRB creating a TP53BP1-PDGFRB fusion protein.
  • Sequence similarities

    Contains 2 BRCT domains.
  • Post-translational
    modifications

    Asymmetrically dimethylated on Arg residues by PRMT1. Methylation is required for DNA binding.
    Phosphorylated at basal level in the absence of DNA damage. Hyper-phosphorylated in an ATM-dependent manner in response to DNA damage induced by ionizing radiation. Hyper-phosphorylated in an ATR-dependent manner in response to DNA damage induced by UV irradiation.
  • Cellular localization

    Nucleus. Chromosome > centromere > kinetochore. Associated with kinetochores. Both nuclear and cytoplasmic in some cells. Recruited to sites of DNA damage, such as double stand breaks. Methylation of histone H4 at 'Lys-20' is required for efficient localization to double strand breaks.
  • Information by UniProt
  • Database links

  • Alternative names

    • 53 BP1 antibody
    • 53BP1 antibody
    • FLJ41424 antibody
    • MGC138366 antibody
    • p202 antibody
    • p53 binding protein 1 antibody
    • p53 BP1 antibody
    • p53-binding protein 1 antibody
    • p53BP1 antibody
    • TP53 BP1 antibody
    • TP53B_HUMAN antibody
    • Tp53bp1 antibody
    • TRP53 BP1 antibody
    • Tumor protein 53 binding protein 1 antibody
    • Tumor protein p53 binding protein 1 antibody
    • Tumor suppressor p53 binding protein 1 antibody
    • Tumor suppressor p53-binding protein 1 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling 53BP1 with ab21083 at 1/4000 dilution. Nuclear staining is observed. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

  • All lanes : Anti-53BP1 antibody (ab21083) at 1/500 dilution

    Lane 1 : 53BP1 shRNA non-transfected HeLa whole cell extracts
    Lane 2 : 53BP1 shRNA transfected HeLa whole cell extracts

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG antibody

    Predicted band size: 220 kDa
    Observed band size: 350 kDa
    why is the actual band size different from the predicted?



    5% SDS-PAGE

  • All lanes : Anti-53BP1 antibody (ab21083) at 1/2000 dilution

    Lane 1 : 293T whole cell lysate/extract
    Lane 2 : A431 whole cell lysate/extract
    Lane 3 : HeLa whole cell lysate/extract
    Lane 4 : HepG2 whole cell lysate/extract
    Lane 5 : A375 whole cell lysate/extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG antibody

    Predicted band size: 220 kDa
    Observed band size: 350 kDa why is the actual band size different from the predicted?



    5% SDS-PAGE.

    Running conditions: 80V, 15min; 140V, 40min.

    Transfer condition: Semi-dry, 18 V, 60min (Nitrocellulose membrane).

    Blocking condition: 5% non-fat milk in TBST, RT, 60min.

    Primary antibody incubation: 1/2000, 4?, overnight.

    Secondary antibody incubation: Rabbit IgG antibody (HRP), 1/10,000, RT, 1hr. 

    Washing condition: 5 ml TBST, 4 x 5min.

    Exposure system: Trident plus Western HRP Substrate.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling 53BP1 with ab21083 at 1/4000 dilution. Nuclear staining is observed. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

  • All lanes : Anti-53BP1 antibody (ab21083) at 1/500 dilution

    Lane 1 : Neuro2A whole cell extracts
    Lane 2 : C8D30 whole cell extracts
    Lane 3 : NIH-3T3 whole cell extracts

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG antibody

    Predicted band size: 220 kDa
    Observed band size: 350 kDa why is the actual band size different from the predicted?



    5% SDS-PAGE

  • ab21083 (2µg/ml) staining 53BP1 in human Brain: Cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is ubiquitous nuclear staining throughout.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

References

This product has been referenced in:

  • Riaz MA  et al. Metformin enhances the radiosensitizing effect of cisplatin in non-small cell lung cancer cell lines with different cisplatin sensitivities. Sci Rep 9:1282 (2019). Read more (PubMed: 30718758) »
  • Guard SE  et al. The nuclear interactome of DYRK1A reveals a functional role in DNA damage repair. Sci Rep 9:6539 (2019). Read more (PubMed: 31024071) »
See all 61 Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A549 (NSCLC))
Permeabilization
No
Specification
A549 (NSCLC)
Blocking step
10% FCS + 0.2% Triton X-100 as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Methanol

Simon Deycmar

Verified customer

Submitted Feb 21 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (head and neck cancer)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (8)
Loading amount
15 µg
Treatment
drug 24h
Specification
head and neck cancer
Blocking step
BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 06 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (MRC5VA)
Loading amount
15 µg
Specification
MRC5VA
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Apr 19 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (T lymphocyte)
Specification
T lymphocyte
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% triton
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Nov 29 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa and SK-N-SH)
Specification
HeLa and SK-N-SH
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jul 12 2006

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