Nbs1 Antibody (NB100-143)

Images

 
Western Blot: Nbs1 Antibody [NB100-143] - TIG-1 human primary fibroblasts, whole cell lysate (30 ug). Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody [NB100-143] - Hela cells. Image from verified customer review.
Immunohistochemistry-Paraffin: Nbs1 Antibody [NB100-143] - IHC analysis of formalin fixed paraffin-embedded (FFPE) human testis using Nbs1 antibody at 1:200 on a Bond Rx autostainer (Leica Biosystems). The assay ...read more
Western Blot: Nbs1 Antibody [NB100-143] - WB analysis of HeLa whole cell lysate (NB800-PC1) using rabbit polyclonal NBS1 antibody (NB100-143) with detection employing ECL method.
Western Blot: Nbs1 Antibody [NB100-143] - Detection of NBS1 in Human Bone Osteosarcoma Epithelial Cells (U2OS whole cell lysate) using NB100-143 at a dilution of 1/1000. Specificity was confirmed using a siRNA anti-NBS1 ...read more
Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody [NB100-143] - Staining of Hela cells using NB100-143 anti-NBS1.

Product Details

Summary
Reactivity Hu, Mu, Ha, Ma, PmSpecies Glossary
Applications WB, ChIP, Flow, ICC/IF, IHC, IHC-P, IP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
     

Genetic Strategies

   

Order Details

Nbs1 Antibody Summary

Immunogen
Partial length human NBS1 protein [Swiss-Prot# O60934].
Localization
Nuclear
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
NBN
Purity
Unpurified
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
  • Chromatin Immunoprecipitation
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunohistochemistry 1:100-1:200
  • Immunohistochemistry-Paraffin 1:100 - 1:200
  • Immunoprecipitation 3 ul
Application Notes
This NBS1 antibody is useful for Western blot, Immunoprecipitation and Immunocytochemistry/Immunofluorescence. Use in Chromatin Immunoprecipitation was reported in scientific literature (PMID: 23681918). By WB, this NBS1 antibody recognizes a band at 95 kDa, representing NBS1. By ICC/IF, this antibody has been used with methanol-fixed IMR90 primary human fibroblasts. IP tests have been done with 3-4 X 106 cells. 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.
Theoretical MW
95 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
Nbs1 Lysate (NBL1-13497)
Reviewed Applications
Read 8 Reviews rated 4.8
using
NB100-143 in the following applications:

Publications
Read Publications using
NB100-143 in the following applications:

  • 4 publications
  • 3 publications
  • 38 publications
  • IHC
    4 publications
  • IP
    16 publications
  • WB
    52 publications

Reactivity Notes

Mammal reactivity reported in scientific literature (PMID: 24064949)

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
Whole antisera
Preservative
0.02% Sodium Azide
Purity
Unpurified

Alternate Names for Nbs1 Antibody

  • ATV
  • AT-V1
  • AT-V2
  • Cell cycle regulatory protein p95
  • FLJ10155
  • MGC87362
  • NBN
  • Nbs1
  • NBS1P95
  • NBSNijmegen breakage syndrome protein 1
  • Nibrin
  • Nijmegen breakage syndrome 1 (nibrin)
  • p95 protein of the MRE11/RAD50 complex
  • p95

Background

NBS1 (Nijmegen breakage syndrome protein 1) is a component of MRN complex (Mre11-Rad50-Nbs1) that plays important role in detection and signaling of DNA double strand breaks (DSBs) through acting as DSB sensor, co-activator of DSB-induced cell cycle checkpoint signaling, and as repair-effector in two competing DSB repair pathways: homologous recombination (HR) and non-homologous end-joining (NHEJ). MRN complex also associates with telomeres at the ends of linear chromosomes, where it contributes to their maintenance. NBS1's FHA domain binds phospho-Thr residues in Ser-X-Thr motifs present in DNA damage proteins, including Mdc1 and Ctp1, whereas, BRCT domains of NBS1 bind Ser-X-Thr motifs when Ser residue is phosphorylated. These phospho-dependent interactions are important for recruiting repair and checkpoint proteins to DSB sites and NBS1 is responsible for MRN complex's nuclear translocation. NBS1 itself does not possess enzymatic activity and contributes to DSB repair primarily by mediating protein-protein interactions at DNA breakage sites. The central region of NBS1 1 possesses several SQ motifs that are phosphorylated by ATM kinase via DNA damage response. NBS1's C-terminus contains a domain that interacts with ATM and recruits it to DSBs, and induces apoptosis in response to damage. DSBs can be caused by ionizing radiation, certain chemotherapy drugs, metabolic ROS, as errors during replication, by programmed enzymatic activities during meiosis/V(D)J recombination etc., and if left unrepaired, DSBs can generate chromosomal translocations, aneuploidy and carcinogenesis.

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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Species: Hu, Mu, Ha, Ma, Pm
Applications: WB, ChIP, Flow, ICC/IF, IHC, IHC-P, IP

Publications for Nbs1 Antibody (NB100-143)(154)

We have publications tested in 6 confirmed species: Human, Mouse, Ape, Hamster, Mammal, Primate.

We have publications tested in 6 applications: ChIP, FLOW, ICC/IF, IHC, IP, WB.


Filter By Application
ChIP
(4)
FLOW
(3)
ICC/IF
(38)
IHC
(4)
IP
(16)
WB
(52)
All Applications
Filter By Species
Human
(74)
Mouse
(7)
Ape
(1)
Hamster
(1)
Mammal
(1)
Primate
(1)
All Species
Showing Publications 1 - 10 of 154. Show All 154 Publications.
Publications using NB100-143 Applications Species
Ward TA, McHugh PJ, Durant ST. Small molecule inhibitors uncover synthetic genetic interactions of human flap endonuclease 1 (FEN1) with DNA damage response genes. PLoS ONE. 2017 Jun 19 [PMID: 28628639] (WB, Human) WB Human
Burke RT, Marcus JM, Orth JD. Inhibition of exportin-1 function results in rapid cell cycle-associated DNA damage in cancer cells Oncotarget 2017 Jun 13 [PMID: 28467801] (ICC/IF, Human) ICC/IF Human
Mariggio G, Koch S, Zhang G et al. Kaposi Sarcoma Herpesvirus (KSHV) Latency-Associated Nuclear Antigen (LANA) recruits components of the MRN (Mre11-Rad50-NBS1) repair complex to modulate an innate immune signaling pathway and viral latency. PLoS Pathog. Apr 21 2017 [PMID: 28430817]
Smith EA, Gole B, Willis NA et al. DEK is required for homologous recombination repair of DNA breaks. Sci Rep. Mar 20 2017 [PMID: 28317934] (WB, Human) WB Human
Campos Leon K, Wijendra K, Siddiqa A et al. Association of human papillomavirus type 16 E2 with Rad50-interacting protein 1 enhances viral DNA replication. J. Virol. Dec 28 2016 [PMID: 28031358] (ICC/IF, Human) ICC/IF Human
Lou DI, Kim ET, Meyerson NR et al. An Intrinsically Disordered Region of the DNA Repair Protein Nbs1 Is a Species-Specific Barrier to Herpes Simplex Virus 1 in Primates. Cell Host Microbe. Aug 10 2016 [PMID: 27512903] (WB, Human, Ape) WB Human, Ape
Xu Z, Song Z, Li G et al. H2B ubiquitination regulates meiotic recombination by promoting chromatin relaxation. Nucleic Acids Res 2016 Jul 18 [PMID: 27431324]
Mlody B... Adjaye J, Generation of iPSC lines from a Nijmegen Breakage Syndrome patient. Stem Cell Res 2015 Nov [PMID: 26987925]
Francia S, Cabrini M, Matti V et al. DICER, DROSHA and DNA damage-response RNAs are necessary for the secondary recruitment of DNA damage response factors J. Cell. Sci. Feb 16 2016 [PMID: 26906421] (ICC/IF, Human) ICC/IF Human
Xing M, Wang X, Palmai-Pallag T et al. Acute MUS81 depletion leads to replication fork slowing and a constitutive DNA damage response. Oncotarget 2015 Nov 10 [PMID: 26415217]
Show All 154 Publications.

Reviews for Nbs1 Antibody (NB100-143) (8) 4.88

Average Rating: 4.8
(Based on 8 reviews)
We have 8 reviews tested in 2 species: Human, Other.
We have 8 reviews tested in 2 application: IF, WB.

Reviews using NB100-143:
Filter by Applications
IF
(5)
WB
(3)
All Applications
Filter by Species
Human
(5)
Other
(2)
All Species
Images Ratings Applications Species Date Details
 
reviewed by:
Anonymous
IF Other 12/12/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedSee PMID 24344331
SpeciesOther

Blocking

Blocking DetailsSee PMID 24344331

Details

Detection NotesSee PMID 24344331

Comments

CommentsPublished in PMID: 24344331
 
reviewed by:
Anonymous
IF Human 12/12/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedSee PMID 22013059
SpeciesHuman

Blocking

Blocking DetailsSee PMID 22013059

Details

Detection NotesSee PMID 22013059

Comments

CommentsPublished in PMID: 22013059
 
reviewed by:
Barbara Alcaraz Silva
IF Other 10/08/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedMammalian
SpeciesOther
LotN-2

Primary Anitbody

Dilution Ratio1:500

Details

Fixation Details4% PFA, RT
Immunofluorescence Nbs1 NB100-143
Enlarge
reviewed by:
Svetlana Khoronenkova
IF Human 07/31/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedHeLa cells
SpeciesHuman
LotN-2

Blocking

Blocking Details2% BSA in PBS, 1 h at RT

Primary Anitbody

Dilution Ratio1:1000 in 2% BSA/PBS, 14h at 4C

Secondary Antibody

Secondary DescriptionAlexa Fluor® 488 Goat Anti-Rabbit IgG (H+L)
Secondary Manufacturer Cat#A-11008 from Life Technologies (by Molecular Probes)
Secondary Concentration0.2 ug/ml

Details

Fixation Details4% Paraformaldehyde as fixative, permeabilization with 0.2% Triton-X100
Wash Description3x washes in PBS for 5 min
Western Blot Nbs1 NB100-143
Enlarge
reviewed by:
Svetlana Khoronenkova
WB Human 04/10/2014
View

Summary

ApplicationWestern Blot
Sample TestedTIG-1 human primary fibroblasts, whole cell lysate (30 ug)
SpeciesHuman
LotN-2
CommentsNbs1 knockdown is hard to achieve therefore Mre11 depletion that is known to result in deregulation of Nbs1 levels was used as an additional negative control

Blocking

Blocking DetailsLI-COR® Odyssey® Blocking Buffer for 45 min at RT

Primary Anitbody

Dilution Ratio1:2000 o/n at RT in LI-COR® Odyssey® Blocking Buffer 1:1 in PBS + 0.1%Tween

Secondary Antibody

Secondary DescriptionAlexa Fluor® 680 Goat Anti-Rabbit IgG (H+L)
Secondary Manufacturer Cat#A-21076 from Life Technologies (by Molecular Probes)
Secondary Concentration0.2 ug/ml

Details

Detection NotesLI-COR® Odyssey® detection system, Nbs1 & Mre11 siRNA as a negative control

Comments

CommentsNbs1 knockdown is hard to achieve therefore Mre11 depletion that is known to result in deregulation of Nbs1 levels was used as an additional negative control
Western Blot Nbs1 NB100-143
Enlarge
reviewed by:
Anonymous
WB Human 09/19/2013
View

Summary

ApplicationWestern Blot
Sample TestedU2OS whole cell lysate
SpeciesHuman
CommentsThe antibody performed very well with only few unspecific bands and a specific signal around 100 kDa. Specificity was confirmed using a siRNA anti-NBS1 (targeting sequence: GGAAGAAACGUGAACUCAA).

Blocking

Blocking DetailsMilk

Primary Anitbody

Dilution Ratio1/1000

Secondary Antibody

Secondary DescriptionLI-COR IRDye Donkey anti-Rabbit 800CW
Secondary Manufacturer Cat#926-32213
Secondary Concentration1/15000

Details

Detection NotesOdyssey LI-COR Biosciences

Comments

CommentsThe antibody performed very well with only few unspecific bands and a specific signal around 100 kDa. Specificity was confirmed using a siRNA anti-NBS1 (targeting sequence: GGAAGAAACGUGAACUCAA).
Immunofluorescence Nbs1 NB100-143
Enlarge
reviewed by:
Anonymous
IF Human 09/19/2013
View

Summary

ApplicationImmunofluorescence
SpeciesHuman
Special ApplicationsUsed in immunofluorescence in human U2OS cells fixed with paraformaldehyde without pre-extraction or pre-extracted as described in publication PMID:23897892. Gave a signal that is lost after knock-down of NBS1 using siRNA.
 
reviewed by:
Anonymous
WB 01/25/2013
View

Summary

ApplicationWestern Blot
LotJ5
Special ApplicationsThis antibody recognized a Vero cell proteins (Chlorocebus aethiops) that were not tested before. Click <a href="http://www.sciencedirect.com/science/article/pii/S0378113513000564"> here</a> to view article abstract and for more information (in press).

Product General Protocols

View specific protocols for Nbs1 Antibody (NB100-143): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

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

FAQs for Nbs1 Antibody (NB100-143). (Showing 1 - 3 of 3 FAQs).

  1. I would like to use the NBN Antibody for IP. What kind of Ig subclass for NBN is present in the whole antisera (IgG1 etc.) ?
    • As this antibody was produced in rabbit, it is IgG as this is all they produce.
  2. I would like to know if the antibody NB100-143 is suitable to recognize the N-terminal end of the NBS1 protein?
    • Unfortunately, the immunogen sequence for NB100-143 is within the second half of the amino acid sequence of (human) NBS1, and therefore I do not think it would be able to recognize the N-terminal end of the protein.
  3. I have a question about your product, NBS1 antibody (NB100-143). Data sheet describes that the immunogen is partial length human NBS1 protein [Swiss-Prot# O60934]. Would you specifically tell me which portion of NBS1 (with the amino acids number) is used as its immunogen?
    • Our NBS1 product NB100-143 was made to a sequence that falls between amino acids 395-754 of human Nibrin (UniProt O60934). The exact sequence is proprietary, but I hope the immunogen range will be sufficient for your needs.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

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Blogs on Nbs1.

The MRE11 Complex and DNA Damage Response
The maintenance of genome stability depends on the DNA damage response (DDR) which is a complex signaling network including cell cycle checkpoints, DNA repair and damage tolerance pathways. The DDR complex has the ability to sense DNA damage and tra...  Read full blog post.

NBS1: The DNA Repair Trigger
NBS1 (Nijmegen breakage syndrome protein 1) is a component of the MRN complex (Mre11-Rad50-Nbs1) that plays important role in detecting DNA double strand breaks (DSBs) and triggering the downstream cascade. DSBs can be caused by ionizing radiation, ch...  Read full blog post.

NBS1: DNA Repair Trigger
NBS1 (Nijmegen breakage syndrome protein 1) is a component of the MRN complex (Mre11-Rad50-Nbs1) that plays important role in detecting DNA double strand breaks (DSBs) and triggering the downstream cascade. DSBs can be caused by ionizing radiation, ch...  Read full blog post.

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Recent Reviews

4.8
5
6
4
2
3
0
2
0
1
0

 
Anonymous
12/12/2014
Application: IF
Species: Other

 
Anonymous
12/12/2014
Application: IF
Species: Human

 
Barbara Alcaraz Silva
10/08/2014
Application: IF
Species: Other

Bioinformatics

Gene Symbol NBN
Entrez
Uniprot