53BP1 Antibody

Images

 
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-904] - Analysis in HeLa cells. Image from verified customer review.
Western Blot: 53BP1 Antibody [NB100-904] - Analysis in untreated U2OS cell lysate.
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-904] - Analysis in HeLa cells with FITC (green). Nuclei and alpha-tubulin were counterstained with Dapi (blue) and Dylight 550 (red).
Immunohistochemistry: 53BP1 Antibody [NB100-904] - Analysis in human renal cancer using DAB with hematoxylin counterstain.
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-904] - Analysis in C33A cells. Image courtesy of an anonymous customer review.

Product Details

Summary
Reactivity Hu, MuSpecies Glossary
Applications WB, ICC/IF, IHC, IHC-P, IP, PLA
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml

53BP1 Antibody Summary

Immunogen
A synthetic peptide corresponding to a portion of human 53BP1 encoded in exon 11, 12 and 19 [UniProt# Q12888]. This antibody is a cocktail hybrid of NB100-304 and NB100-305.
Marker
DNA Double Strand Break Marker
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
TP53BP1
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:1000-1:10000
  • Immunocytochemistry/Immunofluorescence 1:100-1:1000
  • Immunohistochemistry 1:400
  • Immunohistochemistry-Paraffin 1:400
  • Immunoprecipitation
  • Proximity Ligation Assay
Application Notes
This 53BP1 antibody is useful for Immunocytochemistry/Immunofluorescence, Western Blot and Immunohistochemistry paraffin embedded sections. In Western blot a band was observed at ~220 kD. Irradiated lysates demonstrate a gel shift. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. Use in immunprecipitation reported in scientific literature (PMID 25512560). Use in proximity ligation assay reported in scientific literature (PMID 26734725)
Positive Control
Kidney Lysate (NB820-59429)
Salivary Gland Lysate (NB820-59251)
Reviewed Applications
Read 4 Reviews rated 5
using
NB100-904 in the following applications:

Publications
Read Publications using
NB100-904 in the following applications:

  • 32 publications
  • IHC
    1 publication
  • IP
    1 publication
  • PLA
    1 publication
  • WB
    14 publications

Reactivity Notes

Human has been tested in WB, IHC and ICC/IF, mouse has only been tested in ICC/IF.

Packaging, Storage & Formulations

Storage
Store at -20 °C.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1 mg/ml
Purity
Immunogen affinity purified

Alternate Names for 53BP1 Antibody

  • 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

Background

53BP1 (p53 binding protein 1) plays a key role in response to DNA damage, checkpoint signaling during mitosis and enhancing TP53-mediated transcriptional activation. Originally identified as p53's transcriptional enhancing partner, 53BP1 now has been established as a substrate for ATM (ataxia telangiectasia mutated) signaling and that it relocalizes to discrete foci overlapping with gamma H2AX (phosphorylated histone H2AX); demarcating DNA double strand breaks (DSBs) sites following exposure to radiation. 53BP1 functions downstream of gamma H2AX-dependent hierarchy of proteins that collectively establish IRIF (ionizing radiation induced foci) at DSBs; this hierarchy includes Mre11/Rad50/NBS1 (MRN complex), ATM, MDC1, RNF8, RNF168 and HERC2. With the exception of ATM, whose function to generate gamma H2AX may be partially compensated by the activity of DNA-PK (DNA-dependent kinase), all of these proteins are physically and functionally required to recruit 53BP1 to the DSB site. Briefly, this process involves DSB recognition by MRN, ATM activation, gamma H2AX-formation, MDC1-recruitment, MRN-retention (leading to further ATM-activation and gamma H2AX spreading) and RNF8/RNF168/HERC2-mediated histone H2A and H2AX mono and poly-ubiquitination.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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NB100-904
Species: Hu, Mu
Applications: WB, ICC/IF, IHC, IHC-P, IP, PLA

Publications for 53BP1 Antibody (NB100-904)(70)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 5 applications: ICC/IF, IHC, IP, PLA, WB.


Filter By Application
ICC/IF
(32)
IHC
(1)
IP
(1)
PLA
(1)
WB
(14)
All Applications
Filter By Species
Human
(26)
Mouse
(10)
All Species
Showing Publications 1 - 10 of 70. Show All 70 Publications.
Publications using NB100-904 Applications Species
Hassan S, Esch A, Liby T et al. Pathway-enriched gene signature associated with 53BP1 response to PARP inhibition in triple-negative breast cancer. Mol. Cancer Ther. 2017 Sep 27 [PMID: 28958991] (Human) Human
Bao X, Siprashvili Z, Zarnegar BJ et al. CSNK1a1 Regulates PRMT1 to Maintain the Progenitor State in Self-Renewing Somatic Tissue Dev. Cell 2017 Sep 18 [PMID: 28943242] (IHC, Mouse) IHC Mouse
Sivanand S, Rhoades S, Jiang Q et al. Nuclear Acetyl-CoA Production by ACLY Promotes Homologous Recombination Mol. Cell 2017 Jul 20 [PMID: 28689661]
Foskolou IP, Jorgensen C, Leszczynska KB et al. Ribonucleotide Reductase Requires Subunit Switching in Hypoxia to Maintain DNA Replication. Mol. Cell. Apr 20 2017 [PMID: 28416140] (WB, ICC/IF, Human) WB, ICC/IF Human
Ho TT, Warr MR, Adelman ER et al. Autophagy maintains the metabolism and function of young and old stem cells. Nature. Mar 9 2017 [PMID: 28241143]
Li S, Shu FJ, Li Z et al. Cell-type specific role of the RNA-binding protein, NONO, in the DNA double-strand break response in the mouse testes. DNA Repair (Amst) 2017 Mar [PMID: 28209515] (Mouse) Mouse
Kang YJ, Balter B, Csizmadia E et al. Contribution of classical end-joining to PTEN inactivation in p53-mediated glioblastoma formation and drug-resistant survival Nat Commun Jan 17 2017 [PMID: 28094268] (ICC/IF, Mouse) ICC/IF Mouse
Dilley RL, Verma P, Cho NW et al. Break-induced telomere synthesis underlies alternative telomere maintenance. Nature Oct 19 2016 [PMID: 27760120] (WB, Human) WB Human
Moureau S, Luessing J, Harte EC et al. A role for the p53 tumour suppressor in regulating the balance between homologous recombination and non-homologous end joining Open Biol Sep 1 2016 [PMID: 27655732] (ICC/IF, WB, Human) ICC/IF, WB Human
Rantala JK, Kwon S, Korkola J, Gray JW Expanding the Diversity of Imaging-Based RNAi Screen Applications Using Cell Spot Microarrays. Microarrays (Basel) 2013 Apr 11 [PMID: 27605183]
Show All 70 Publications.

Reviews for 53BP1 Antibody (NB100-904) (4) 54

Average Rating: 5
(Based on 4 reviews)
We have 4 reviews tested in 2 species: Human, Mouse.
We have 4 reviews tested in 2 application: ICC, IF.

Reviews using NB100-904:
Filter by Applications
ICC
(1)
IF
(3)
All Applications
Filter by Species
Human
(3)
Mouse
(1)
All Species
Images Ratings Applications Species Date Details
 
reviewed by:
Anonymous
ICC Human 01/20/2017
View

Summary

ApplicationImmunocytochemistry
Sample TestedHeLa cells
SpeciesHuman
Lot012815

Comments

CommentsUsed 1:2000 in 0.5% fish skin gelatin in PBST overnight at 4oC for immunofluorescence. Very good signal to noise ratio.
Immunofluorescence 53BP1 NB100-904
Enlarge
reviewed by:
Anonymous
IF Human 11/20/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedHeLa cells
SpeciesHuman
Lot010714

Blocking

Blocking Details2% BSA in PBS, 1 h at RT

Primary Anitbody

Dilution Ratio1:2000 in 2% BSA/PBS, 14h at 4C

Secondary Antibody

Secondary DescriptionAlexa Fluor® 488 Goat Anti-Mouse IgG (H+L)
Secondary Manufacturer Cat#A-11001 from Life Technologies (by Molecular Probes)
Secondary Concentration0.2 ug/ml

Details

Detection NotesZeiss LSM 780 confocal microscope, x63 oil objective
Fixation Details4% Paraformaldehyde as fixative, permeabilization with 0.2% Triton-X100
Wash Description3x washes in PBS for 5 min
 
reviewed by:
Iris Eke
IF Human 08/24/2012
View

Summary

ApplicationImmunofluorescence
Sample TestedHuman tumor cell lines: A549, FaDu, MiaPaCa2, UTSCC15
SpeciesHuman
Lot041610

Blocking

Blocking Details1% BSA, 30 min, RT

Primary Anitbody

Dilution Ratio1:2000 1h RT

Secondary Antibody

Secondary DescriptionAlexaFluor488 (anti-Rb)
Secondary Manufacturer Cat#Invitrogen A11034
Secondary Concentration1:2000

Details

Detection Notesimages with LSM
Fixation DetailsFixation: 3%FA, Permeabilisation: Triton-X, Buffer: 1%BSA, 1h incubation RT
Wash Description1xPBS, 6x
Immunofluorescence 53BP1 NB100-904
Enlarge
reviewed by:
Olivia Patsalos
IF Mouse 05/22/2012
View

Summary

ApplicationImmunofluorescence
Sample TestedMEFs, HeLa, C33A, CaSki
SpeciesMouse
Lot041912
CommentsComplete protocol can be found at: delangelab.rockefeller.edu/protocols_files/IF_protocol.pdf

Blocking

Blocking DetailsPBG blocking 30mins @RT

Primary Anitbody

Dilution Ratio1:500 2h @RT

Secondary Antibody

Secondary DescriptionFITC conjugated Donkey anti-rabbit IgG
Secondary Manufacturer Cat#711-095-152
Secondary Concentration1:250

Details

Detection NotesZeiss Observer A1, Axiovision
Fixation Details4% PFH fixation 10mins @RT, Triton-X permeabilization 10mins @RT, PBS wash 2x5mins @RT
Wash DescriptionPBG wash 3x5mins @RT

Comments

CommentsComplete protocol can be found at: delangelab.rockefeller.edu/protocols_files/IF_protocol.pdf

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for 53BP1 Antibody (NB100-904). (Showing 1 - 5 of 5 FAQs).

  1. We're looking for anti53BP1 antibody covalently labeled with a fluorofor preferabley Cy3. Do you make it?
    • We sell 53BP1 antibodies available conjugated to DyLight 488, 550 (very similar in spectrum to Cy3) and 650. You may also purchase the unlabeled antibodies and conjugate them to the fluorophores of your choice. The catalog numbers that you may be interested in are: NB100-904R, NB100-304R, NB100-305R.
  2. Hello I want to ask you for an advice in case of antibodies. In our research we focus on DNA repair foci and use your Novus antibodies for phosphorylated histone H2AX (monoclonal mouse) and protein 53BP1(polyclonal rabbit). Now we want to use antobody for phosphorylated 53BP1 protein to increase specifity and I want to ask you which from your NOVUS antibodies will be the best for us.
    • We currently stock just one antibody to phosphorylated 53BP1, which you can see at the following link: NB100-1803. This is guaranteed for detection of 53BP1 (pSer25) in human and mouse samples by WB, FLOW, ICC/IF,IHC-P, IP and PLA. As this antibody is a rabbit polyclonal, you would be unable to stain for both 53BP1 and its phosphorylated isoform in the same sample, unless you used an antibody from a different host for detection of total 53BP1 or used directly conjugated primaries. Our mouse monoclonal to 53BP1, with catalogue number NBP2-25028, is validated for detection of the human protein by WB, FLOW and IHC. Unfortunately we do not currently stock any other antibodies to phosphorylated 53BP1.
  3. I would like to know if these antibodies are suitable with IHC techniques using HRP conjugated antibodies and DAB as chromogen? Cat# NB200-175, NB100-904 and NB100-142
    • Yes, all of these antibodies are useful for IHC-P with DAB based detection. Please note that the PTEN Antibody (28H6) NB200-175 has been discontinued, however.
  4. I would like to know if this ab would also work for phosphorylated 53BP1? Or if you have another ab that would detect it?
    • NB100-904 is a cocktail of NB100-304 and NB100-305. These two antibodies recognize between residues 350 and 400 and residues 1925 and the C-terminus (residue 1972), respectively. There are multiple phosphorylations between residues 350 and 400 and as you can see from our datasheet, NB100-304 recognizes 53BP1. We do sell one phosphospecific antibody to 53BP1, catalog number NB100-1803.
  5. Is there a recommended IHC-P protocol for the 53BP1 Antibody (NB100-904)?
    • We do not have a specific protocol outside of our general protocol for IHC for NB100-904. However, I would ensure that the protocol used includes ample permeabilization for a nuclear signal (usually 0.1-0.5% Triton X in the blocking and diluent buffers or a separate permeabilization step) and heat induced antigen retrieval with pH 6.0 citrate buffer. Our general protocols for both IHC-P and antigen retrieval are linked below. IHC-P Protocol Antigen Retrieival Protocol

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB100-904AF405
Alexa Fluor 488 Labeled NB100-904AF488
Alexa Fluor 647 Labeled NB100-904AF647
Alexa Fluor 700 Labeled NB100-904AF700
Biotin Labeled NB100-904B
DyLight 405 Labeled NB100-904V
DyLight 488 Labeled NB100-904G
DyLight 550 Labeled NB100-904R
DyLight 650 Labeled NB100-904C
FITC Labeled NB100-904F
HRP Labeled NB100-904H

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Blogs on 53BP1.

The recent relationship of BRCA1 and 53BP1
The p53-binding protein 1 (53BP1) is a DNA damage response factor, which is recruited to nuclear structures at the site of DNA damage.  DNA double-strand breaks (DSBs) are mutations that are detrimental to cell viability and genome stability, and m...  Read full blog post.

53BP1 - a marker for DNA Double Strand Break
53BP1 (p53 binding protein 1) was originally thought to be an enhancer for p53 transcriptional, but later studies have demonstrated that it is actually a substrate for ataxia telangiectasia mutated (ATM). 53BP1 is a classic late DNA damage response...  Read full blog post.

53BP1 - DNA damage is no fun
The 53BP1 (p53 binding protein 1) was initially believed to be a p53 transcriptional enhancing partner, but it has now been established as an ataxia telangiectasia mutated (ATM) substrate. As a late DNA damage response (DDR) marker, 53BP1 appears duri...  Read full blog post.

53BP1, DNA Damage Response and Tumor Suppression
53BP1 (p53 binding protein 1) was originally thought to be a p53 transcriptional enhancing partner, but now has been shown to be an ataxia telangiectasia mutated (ATM) substrate. It is a late DNA damage response (DDR) marker, appearing in the telophas...  Read full blog post.

53BP1, DNA Damage Response and Tumor Suppression
53BP1 (p53 binding protein 1) was originally thought to be a p53 transcriptional enhancing partner, but now has been shown to be an ataxia telangiectasia mutated (ATM) substrate. It is a late DNA damage response (DDR) marker, appearing in the telophas...  Read full blog post.

NUP153 & 53BP1: A Novel DNA Repair Pathway
Mediating DNA damage is a crucial process, and one of the most important cellular guards against cancer. In response to DNA damage, sophisticated cellular machinery is recruited to repair the breaks, and if it fails, the cell is committed to death. De...  Read full blog post.

Blocking 53BP1 Expression Lessens Tumor Development in BRCA1-Defective Mice
Our antibody database at Novus Biologicals provides research tools for the forefront of cancer research. Recently, a mouse study using 53BP1 and BRCA1 antibodies showed that deletion of 53BP1 greatly lessened the incidence of tumor development in mice...  Read full blog post.

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Recent Reviews

5
5
4
4
0
3
0
2
0
1
0

 
Anonymous
01/20/2017
Application: ICC
Species: Human

Anonymous
11/20/2014
Application: IF
Species: Human

 
Iris Eke
08/24/2012
Application: IF
Species: Human

Bioinformatics

Gene Symbol TP53BP1
Entrez
Uniprot