53BP1 Antibody

Images

 
Immunohistochemistry-Frozen: 53BP1 Antibody [NB100-304] - Irradiated cochlear spiral ganglion cells, mouse, frozen section of fixed material. Data and image from confirmed customer review.
Western Blot: 53BP1 Antibody [NB100-304] - Detection of 53BP1 by Western Blot Sample: Whole cell lysate from U2OS or 293T cells.
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.
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] - 53BP1 antibody was tested at 1:100 in HeLa cells with FITC (green). alpha-Tubulin antibody NB100-690 was tested at a 1:500 dilution using Mouse IgG ...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
Immunohistochemistry-Paraffin: 53BP1 Antibody [NB100-304] - IHC staining of 53BP1 in human colon cancer using DAB with hematoxylin counterstain.

Product Details

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

Biological Strategies

     

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
  • Knockout Validated
Application Notes
This 53BP1 antibody is useful for Western Blot, Immunohistochemistry on paraffin-embedded sections, Immunocytochemistry and Flow Cytometry. 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.
Positive Control
Rectum Lysate (NB820-59448)
Ovary Lysate (NB820-59442)
Reviewed Applications
Read 8 Reviews rated 4.4
using
NB100-304 in the following applications:

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

Reactivity Notes

Human and mouse 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
  • 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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NB100-304
Species: Hu, Mu, Rt, Fi, Gt, Pm
Applications: WB, ChIP, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KO

Publications for 53BP1 Antibody (NB100-304)(282)

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
(169)
IHC
(12)
IHC-Fr
(2)
IHC-P
(13)
IP
(3)
WB
(38)
All Applications
Filter By Species
Human
(133)
Mouse
(74)
Rat
(3)
Goat
(1)
Primate
(1)
All Species
Showing Publications 1 - 10 of 282. Show All 282 Publications.
Publications using NB100-304 Applications Species
Jones SE, Fleuren EDG, Frankum J et al. ATR is a therapeutic target in synovial sarcoma. Cancer Res. 2017 Oct 16 [PMID: 29038346] (ICC/IF, Human) ICC/IF Human
Hosokawa K, MacArthur BD, Ikushima YM et al. The telomere binding protein Pot1 maintains haematopoietic stem cell activity with age. Nat Commun. 2017 Oct 06 [PMID: 28986560] (Flow, Mouse) Flow Mouse
Schuhwerk H, Bruhn C, Siniuk K et al. Kinetics of poly (ADP-ribosyl) ation, but not PARP1 itself, determines the cell fate in response to DNA damage in vitro and in vivo Nucleic Acids Research 2017 Aug 18 [PMID: 28977496] (Mouse) Mouse
Patel DS, Misenko SM, Her J, Bunting SF. BLM helicase regulates DNA repair by counteracting RAD51 loading at DNA double-strand break sites J. Cell Biol. 2017 Sep 14 [PMID: 28912125] (Human) Human
Traver G, Mont S, Gius D et al. Loss of Nrf2 promotes alveolar type 2 cell loss in irradiated, fibrotic lung Free Radic. Biol. Med. 2017 Nov 01 [PMID: 28870520] (WB, Mouse) WB Mouse
Perkhofer L, Schmitt A, Romero Carrasco MC et al. ATM deficiency generating genomic instability sensitizes pancreatic ductal adenocarcinoma cells to therapy-induced DNA damage Cancer Res. 2017 Aug 08 [PMID: 28790064] (Human) Human
Lai X, Broderick R, Bergoglio V et al. MUS81 nuclease activity is essential for replication stress tolerance and chromosome segregation in BRCA2-deficient cells Nat Commun 2017 Jul 17 [PMID: 28714477] (ICC/IF, Human) ICC/IF Human
Mikula-Pietrasik J, Uruski P, Pakula M et al. Oxidative stress contributes to hepatocyte growth factor-dependent pro-senescence activity of ovarian cancer cells. Free Radic. Biol. Med. 2017 Jun 23 [PMID: 28652056]
Liu R, King A, Hoch NC et al. ASCIZ/ATMIN is dispensable for ATM signaling in response to replication stress. DNA Repair (Amst.). 2017 Jun 17 [PMID: 28648892] (WB, ICC/IF, Mouse) WB, ICC/IF Mouse
Yosef R, Pilpel N, Papismadov N et al. p21 maintains senescent cell viability under persistent DNA damage response by restraining JNK and caspase signaling. EMBO J. 2017 Jun 12 [PMID: 28607003] (ICC/IF, Human) ICC/IF Human
Show All 282 Publications.

Reviews for 53BP1 Antibody (NB100-304) (8) 4.48

Average Rating: 4.4
(Based on 8 reviews)
We have 8 reviews tested in 3 species: Human, Mouse, Human and Mouse.
We have 8 reviews tested in 5 application: ICC, IF, WB, IHC-Fr, IHC.

Reviews using NB100-304:
Filter by Applications
ICC
(4)
IF
(1)
WB
(1)
IHC-Fr
(1)
IHC
(1)
All Applications
Filter by Species
Human
(3)
Mouse
(4)
Human and Mouse
(1)
All Species
Images Ratings Applications Species Date Details
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
 
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
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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
Enlarge
reviewed by:
leslie liberman
IHC-Fr Mouse 01/21/2014
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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 Video Protocol
ChIP Webinar
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.

Positive 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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Bioinformatics Tool for 53BP1 Antibody (NB100-304)

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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

4.4
5
3
4
5
3
0
2
0
1
0

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

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

 
Anonymous
05/22/2017
Application: ICC
Species: Human

Bioinformatics

Gene Symbol TP53BP1
Entrez