53BP1 Antibody

Images

 
Western Blot: 53BP1 Antibody [NB100-304] - Total protein from HeLa, A431, Neuro2A, and PC12 was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was ...read more
Genetic Strategies: Knockout Validated: 53BP1 Antibody [NB100-304] - 53BP1 was detected in immersion fixed HeLa cells (left) but was not detected in 53BP1 knockout Hela cells (right) using Rabbit Anti-human 53BP1 ...read more
Biological Strategies: Immunohistochemistry-Frozen: 53BP1 Antibody [NB100-304] - Irradiated cochlear spiral ganglion cells, mouse, frozen section of fixed material. Image from verified customer review.
Biological Strategies: Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - 53BP1 foci in proliferating MEFs; both normal and exposed to 10 Gy of IR.
Immunohistochemistry-Paraffin: 53BP1 Antibody [NB100-304] - Detection of Human and Mouse 53BP1 by IHC. Sample: FFPE sections of human ovarian carcinoma (left) and mouse teratoma (right). Antibody: NB100-304 used at a ...read more
Flow Cytometry: 53BP1 Antibody [NB100-304] - 1 million Jurkat cells were fixed, permeabilized, and stained with 1.5 ug/ml anti-53BP1 NB100-304 in a 150 uL reaction. Isotype control (black), anti-53BP1 (red).
Immunohistochemistry-Paraffin: 53BP1 Antibody [NB100-304] - Human breast tumors stained with 53BP1 antibody. Image from verified customer review.
Western Blot: 53BP1 Antibody [NB100-304] - Whole cell lysate from U2OS or 293T cells.
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - PC12 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were ...read more
Flow Cytometry: 53BP1 Antibody [NB100-304] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody NB100-304AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - 53BP1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using 1 ug/mL of rabbit anti- 53BP1 polyclonal (NB100-304, ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - Neuro2a cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - Human medulloblastoma (DAOY) and mouse astrocyte (C8D1A) cell lines were exposed for 48 hours to DMSO or 1ug/mL of the DNA damaging agent ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - Embryonic Fibroblast cells pre-extraction for 5 mins with CSK buffer. Fixed with 4% PFA and 75% Ethanol. Primary Antibody at 1:1000. Secondary ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - Upper Panel: Control untreated cells. Lower Panel: Cells exposed to irradiation (10 Gy) and probed for 53BP1 foci. Cells were grown on coverslips, ...read more
Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody [NB100-304] - NIH3T3 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were ...read more
Immunohistochemistry-Paraffin: 53BP1 Antibody [NB100-304] - Staining of 53BP1 in human colon cancer using DAB with hematoxylin counterstain.
Flow (Intracellular): 53BP1 Antibody [NB100-304] - An intracellular stain was performed on Neuro2A cells with NB100-304F (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then ...read more
Flow (Intracellular): 53BP1 Antibody [NB100-304] - An intracellular stain was performed on Neuro2A cells with NB100-304G (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Fi, Gt, PmSpecies Glossary
Applications WB, ChIP, Flow, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, IP, ICC/IF, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml

53BP1 Antibody Summary

Immunogen
The epitope recognized by this antibody maps to a region between residues 350 and 400 of human 53BP1 [NP_005648.1].
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 1:5000 - 1:25000
  • Chromatin Immunoprecipitation
  • Flow Cytometry 1.5 ug/mL
  • Immunocytochemistry/Immunofluorescence 1:1000 - 1:5000
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:1000 - 1:5000
  • Immunoprecipitation
  • Flow (Intracellular)
  • Immunocytochemistry 1:200
  • Knockout Validated
Application Notes
Epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections. Immunohistochemistry-Paraffin and Immunohistochemistry were reported in scientific literature. Frozen section data from customer review. Use in chromatin immunoprecipitation reported in scientific literature (PMID 24591601) Use in Immunoprecipitation reported in scientific literature (PMID 25645366). Knock Out Validation was reported in scientific literature (PMID: 26601238).
Theoretical MW
214 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Control
Rectum Lysate (NB820-59448)
Ovary Lysate (NB820-59442)
Reviewed Applications
Read 10 Reviews rated 4.5
using
NB100-304 in the following applications:

Publications
Read Publications using
NB100-304 in the following applications:

Reactivity Notes

Human and mouse reactivity cited in numerous publications. Primate reactivity reported in scientific literature (PMID: 18389475). Fish reactivity reported in scientific literature (PMID: 25516420).

Packaging, Storage & Formulations

Storage
Store at -20 °C.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1 mg/ml
Purity
Immunogen affinity purified

Notes

Please note for Lot A5, the dilution recommendations have changed from our previous lot recommendations. You may be required to use the antibody at a higher dilution than previous lots to obtain similar results. Please contact us if you have any questions.

Alternate Names for 53BP1 Antibody

  • 53BP1
  • FLJ41424
  • MGC138366
  • p202
  • p53-binding protein 1
  • p53bp1
  • 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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Publications for 53BP1 Antibody (NB100-304)(327)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Goat, Primate.

We have publications tested in 11 applications: ChIP, FISH, FLOW, Flow, ICC-IF, ICC/IF, IHC, IHC-Fr, IHC-P, IP, WB.


Filter By Application
ChIP
(3)
FISH
(1)
FLOW
(4)
Flow
(2)
ICC-IF
(1)
ICC/IF
(196)
IHC
(12)
IHC-Fr
(2)
IHC-P
(14)
IP
(3)
WB
(47)
All Applications
Filter By Species
Human
(164)
Mouse
(82)
Rat
(3)
Goat
(1)
Primate
(1)
All Species
Showing Publications 1 - 10 of 327. Show All 327 Publications.
Publications using NB100-304 Applications Species
Munoz-Galvan S, Lucena-Cacace A, Perez M et al. Tumor cell-secreted PLD increases tumor stemness by senescence-mediated communication with microenvironment. Oncogene. Oct 10 2018 [PMID: 30305726] (ICC/IF, Human) ICC/IF Human
Tete A, Gallais I, Imran M et al. Mechanisms involved in the death of steatotic WIF-B9 hepatocytes co-exposed to benzo[a]pyrene and ethanol: a possible key role for xenobiotic metabolism and nitric oxide. Free Radic. Biol. Med. Sep 27 2018 [PMID: 30268890] (ICC/IF, Human) ICC/IF Human
Albers E, Sbroggio M, Pladevall-Morera D et al. Loss of PICH Results in Chromosomal Instability, p53 Activation, and Embryonic Lethality. Cell Rep Sep 18 2018 [PMID: 30232008] (ICC/IF, Mouse) ICC/IF Mouse
Johnson TI, Costa ASH, Ferguson AN, Frezza C. Fumarate hydratase loss promotes mitotic entry in the presence of DNA damage after ionising radiation. Cell Death Dis Sep 6 2018 [PMID: 30190474] (ICC/IF, Human) ICC/IF Human
Chow TT, Shi X, Wei JH et al. Local enrichment of HP1alpha at telomeres alters their structure and regulation of telomere protection. Nat Commun Sep 4 2018 [PMID: 30181605] (ICC/IF, Human) ICC/IF Human
Xia Y, Ivanovska IL, Zhu K et al. Nuclear rupture at sites of high curvature compromises retention of DNA repair factors. J. Cell Biol. Aug 31 2018 [PMID: 30171044] (ICC/IF, Human) ICC/IF Human
Petroni M, Sardina F, Infante P et al. MRE11 inhibition highlights a replication stress-dependent vulnerability of MYCN-driven tumors. Cell Death Dis Aug 30 2018 [PMID: 30166519] (ICC/IF, IHC-P, Human) ICC/IF, IHC-P Human
Hehlgans S, Booms P, Gullulu O et al. Radiation Sensitization of Basal Cell and Head and Neck Squamous Cell Carcinoma by the Hedgehog Pathway Inhibitor Vismodegib Int J Mol Sci Aug 23 2018 [PMID: 30142876] (ICC/IF, Human) ICC/IF Human
Muoio D, Berardinelli F, Leone S et al. Naphthalene diimide-derivatives G-quadruplex ligands induce cell proliferation inhibition, mild telomeric dysfunction and cell cycle perturbation in U251MG glioma cells FEBS J. Aug 10 2018 [PMID: 30095224] (ICC/IF, Human) ICC/IF Human
de Carcer G, Venkateswaran SV, Salgueiro L et al. Plk1 overexpression induces chromosomal instability and suppresses tumor development Nat Commun Aug 1 2018 [PMID: 30069007] (Mouse) Mouse
Show All 327 Publications.

Reviews for 53BP1 Antibody (NB100-304) (10) 4.510

Average Rating: 4.5
(Based on 10 reviews)
We have 10 reviews tested in 3 species: Human, Mouse, Human and Mouse.

Reviews using NB100-304:
Filter by Applications
WB
(2)
ICC
(5)
IF
(1)
IHC-Fr
(1)
IHC
(1)
All Applications
Filter by Species
Human
(4)
Mouse
(5)
Human and Mouse
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot 53BP1 NB100-304
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reviewed by:
Anonymous
WB Mouse 11/27/2018
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Summary

ApplicationWestern Blot
Sample Testedmouse epithelial pancreatic cancer cells
SpeciesMouse
LotD-9 and D-10

Comments

CommentsMembrane was blocked with 5% non-fat dry milk, 1h RT. Then incubated ON 4C with 53bp1 antibody 1:10000. After washes, membrane was incubated with HRP-conjugated secondary antibody diluted in 5% non-fat dry milk.
Immunocytochemistry 53BP1 NB100-304
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reviewed by:
Vincent Picco
ICC Human and Mouse 08/23/2017
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Summary

ApplicationImmunocytochemistry
Sample TestedCultured cell lines
SpeciesHuman and Mouse
LotD-6
Immunofluorescence 53BP1 NB100-304
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reviewed by:
Anonymous
IF Mouse 07/18/2017
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Summary

ApplicationImmunofluorescence
Sample TestedMEF
SpeciesMouse
Immunocytochemistry 53BP1 NB100-304
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reviewed by:
Anonymous
ICC Human 06/26/2017
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Summary

ApplicationImmunocytochemistry
Sample TestedA172 Glioblastoma cells
SpeciesHuman
LotD-2-092616-AF488

Comments

CommentsCells were grown on coverslips, fixed with 4% paraformaldehyde, methanol permeabilized, blocked for 1 h, RT. Incubated with 53BP1 (AF488 conjugated) antibody (1:200) overnight, washed 3x with PBS, probed with tubulin (Alexa fluor 594) antibody for 2 h, RT. Washed 3x with PBS, mounted on slides using prolong gold, imaged using Nikon confocal microscope (100x oil).
 
reviewed by:
Anonymous
ICC Human 05/22/2017
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Summary

ApplicationImmunocytochemistry
Sample Tested hela cell, fibroblasts
SpeciesHuman
Immunocytochemistry 53BP1 NB100-304
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reviewed by:
Anonymous
ICC Human 01/04/2016
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Summary

ApplicationImmunocytochemistry
SpeciesHuman
Lotmultiple lots
Special ApplicationsThis antibody also works for western and immunofluorescence.
FileView PDF
 
reviewed by:
Anonymous
WB Human 12/12/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 23755068
SpeciesHuman

Blocking

Blocking DetailsSee PMID 23755068

Details

Detection NotesSee PMID 23755068

Comments

CommentsPublished in PMID: 23755068
Immunohistochemistry-Frozen 53BP1 NB100-304
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reviewed by:
leslie liberman
IHC-Fr Mouse 01/21/2014
View

Summary

ApplicationImmunohistochemistry-Frozen
Sample Testedmouse cochlear spiral ganglion cells
SpeciesMouse
Lot3
CommentsThis antibody co-localizes with anti-H2AX.

Blocking

Blocking Details5% normal horse serum, one hr @ rm temp

Primary Anitbody

Dilution Ratio1:500 in 1% normal horse serum with 0.3% triton-x, overnight at rm temp

Secondary Antibody

Secondary Descriptionchicken anti-rabbit IgG, AlexaFluor488 conjugate, one hr at rm temp
Secondary Manufacturer Cat#Life Technologies #A21441
Secondary Concentration1:1000, diluted in 1% normal horse serum

Details

Detection NotesImaged on a Leica confocal. Tests on non-irradiated tissue, and secondaries-only incubation, produced no label.
Fixation Details4% paraformaldehyde fixation, no antigen retrieval, permeabilized with 1% triton (in block), one hr @ rm temp
Wash DescriptionPBS, 3 changes, 10 min. total

Comments

CommentsThis antibody co-localizes with anti-H2AX.
 
reviewed by:
Anonymous
IHC Mouse 07/14/2010
View

Summary

ApplicationImmunohistochemistry
Sample TestedMouse thymocytes
SpeciesMouse
CommentsI needed to localize DSB repair at a single locus, so IHC was combined with DNA FISH. Many other antibodies failed, as DNA FISH requires heating the cells to 80 degrees C which denatures most proteins. This anti-53BP1 antibody resulted in discrete, bright foci regardless of FISH hybridization.

Comments

CommentsI needed to localize DSB repair at a single locus, so IHC was combined with DNA FISH. Many other antibodies failed, as DNA FISH requires heating the cells to 80 degrees C which denatures most proteins. This anti-53BP1 antibody resulted in discrete, bright foci regardless of FISH hybridization.
 
reviewed by:
Anonymous
ICC Mouse 05/27/2009
View

Summary

ApplicationImmunocytochemistry
Sample Testedmouse B cell
SpeciesMouse
LotA

Blocking

Blocking DetailsBlocking Buffer: BSA, Blocking Time: 30 minutes, Blocking Temp: room temperature

Primary Anitbody

Dilution RatioIncubation Dilution Buffer: pbs + 0.2% BSA + 0.1% tween 20, Dilution Ratio: 1/500, Incubation Time: 1 hour, Incubation Temp: RT

Secondary Antibody

Secondary DescriptionSecondary Ab: alexa fluor 633 goat anti rabbit, Secondary Ab Dilution: 1/500
Secondary Manufacturer Cat#invitrogen

Product General Protocols

Video Protocols

WB Video Protocol
ChIP Webinar
ChIP Video Protocol
ICC/IF Video Protocol

FAQs for 53BP1 Antibody (NB100-304). (Showing 1 - 4 of 4 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. What is the recommended pH of sodium citrate for the 53bp1 antibody (NB100-304)?
    • I have checked our notes for use of this antibody in IHC, and they say: 1:500 to 1:2,000. Epitope retrieval with citrate buffer pH6.0 is recommended for FFPE tissue sections.
  4. Is possible to know the exact sequence of the epitope used?
    • Unfortunately I am unable to disclose the sequence of this antibody due to proprietary restrictions. At the site recognized by NB100-304, the reference sequences are too divergent to expect NB100-304 to bind to either Bovine or Little Brown Bat. Regarding Mouse Homology, 30/34 sites of the immunogen are homologous Human - 100% homologous. If you would be interested in testing these novel species, please take a look at our Innovator's Reward program.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB100-304AF405
Alexa Fluor 488 Labeled NB100-304AF488
Alexa Fluor 647 Labeled NB100-304AF647
Alexa Fluor 700 Labeled NB100-304AF700
Biotin Labeled NB100-304B
DyLight 405 Labeled NB100-304V
DyLight 488 Labeled NB100-304G
DyLight 550 Labeled NB100-304R
DyLight 650 Labeled NB100-304C
FITC Labeled NB100-304F
HRP Labeled NB100-304H
PE Labeled NB100-304PE

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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

4.5
5
5
4
5
3
0
2
0
1
0

Anonymous
11/27/2018
Application: WB
Species: Mouse

Vincent Picco
08/23/2017
Application: ICC
Species: Human and Mouse

Anonymous
07/18/2017
Application: IF
Species: Mouse

Bioinformatics

Gene Symbol TP53BP1