Nbs1 Antibody

Images

 
Genetic Strategies: Knockout Validated: Nbs1 Antibody [NB100-143] - Lysates of HeLa human cervical epithelial carcinoma parental cell line and Nbs1 knockout (KO) HeLa cell line. PVDF membrane was probed with ...read more
Western Blot: Nbs1 Antibody [NB100-143] - Western Blot with Nbs1 Antibody [NB100-143]. KSHV LANA recruits MRN (Mre11-Rad50-NBS1) complex. Co-immunoprecipitation of endogenous LANA and MRN proteins in BC3 cells. Cells ...read more
Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody [NB100-143] - ICC/IF analysis of Nbs1 in HeLa cells with Nbs1 Antibody [NB100-143]. Image from verified customer review.
Immunohistochemistry-Paraffin: Nbs1 Antibody [NB100-143] - FFPE human testis using Nbs1 antibody [NB100-143] at 1:200 on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen ...read more
Western Blot: Nbs1 Antibody [NB100-143] - Analysis of HeLa whole cell lysate [NB800-PC1] using rabbit polyclonal NBS1 antibody [NB100-143]. Observed Molecular Weight at ~95 kDa.
Genetic Strategies: Western Blot: Nbs1 Antibody [NB100-143] - Detection of NBS1 in Human Bone Osteosarcoma Epithelial Cells (U2OS whole cell lysate) using Nbs1 Antibody [NB100-143] at a dilution of 1:1000. ...read more
Western Blot: Nbs1 Antibody [NB100-143] - Western Blot with 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] - Staining of HeLa cells using Nbs1 Antibody [NB100-143].

Product Details

Summary
Reactivity Hu, Mu, Ha, Ma, PmSpecies Glossary
Applications WB, ChIP, ELISA, Flow, IB, ICC/IF, IHC, IHC-P, IP, ISH, PAGE, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Validated by:
     

Genetic Strategies

   

Order Details

Nbs1 Antibody Summary

Immunogen
Nbs1 Antibody was made to a 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
  • ELISA
  • Flow Cytometry
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunohistochemistry 1:100 - 1:200
  • Immunohistochemistry-Paraffin 1:100 - 1:200
  • Immunoprecipitation 3 ul
  • In-situ Hybridization
  • SDS-Page
  • Knockout Validated
Application Notes
By Western blot, 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 10^6 cells. Use in Chromatin Immunoprecipitation, Flow Cytometry, ICC/IF, Immunoprecipitation, and Western Blot was reported in multiple pieces of scientific literature. Use in In-situ Hybridization reported in scientific literature (PMID: 26415217).
Theoretical MW
85 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
Nbs1 Overexpression Lysate
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
  • 47 publications
  • IHC
    5 publications
  • IP
    18 publications
  • WB
    64 publications

Reactivity Notes

Human and Mouse reactivity reported in mutliple pieces of scientific literature. Sumatran orangutan (Pongo abelii), white-cheeked gibbon (Nomascus leucogenys), and rhesus macaque (Macaca mulatta) reactivity reported in scientific literature (PMID: 27512903). Hamster reactivity reported in scientific literature (PMID: 23255801). Potorous tridactylus reactivity reported in scientific literature (PMID: 24064949). African green monkey 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
  • NBS
  • Nbs1
  • NBS1P95
  • Nibrin
  • Nijmegen breakage syndrome 1 (nibrin)
  • Nijmegen breakage syndrome protein 1
  • p95 protein of the MRE11/RAD50 complex
  • p95

Background

NBS1 (Nijmegen breakage syndrome protein 1, Nibrin) is the eukaryotic component of MRN complex (Mre11-Rad50-Nbs1) involved in homologous recombination repair for DNA double-strand breaks (DSBs) and DNA damage-induced checkpoint activation. NBS1, a nuclear protein with a theoretical molecular weight of 65-85 kDa, is composed of 2 functional regions: the forkhead associated (FHA) N-terminal domain (24-83 amino acids) and the breast cancer C-terminus (BRCT) domain (105-181 amino acids). Mutations in the NBN gene are associated with Nijmegen breakage syndrome, characterized by microcephaly, growth retardation, immunodeficiency, and an increased susceptibility to prostate cancer, lung cancer, liver cancer and intrahepatic cholangiocarcinoma (IHC) (1).

In DNA double strand break repair, the FHA/BRCT domains bind DNA damage sensor proteins including gamma H2AX (phospho Ser139), phosphorylated MDC1 and CtIP (CtBP-interacting protein). NBS1 then recruits the other members of the MRN complex, Mre11 and Rad50, to the proximity of DNA DSBs. The MRN complex has also been shown to interact with ATM or ATR kinases in the presence of DSBs or replication fork stalling, respectively. Phosphorylation of NBS1 at Ser 278 and Ser343 in response to ionizing radiation (IR) is dependent on ATM and is important for activation of the S phase checkpoint. The activation of ATM in response to DNA damage is also facilitated by MRN and may lead to induction of apoptosis (2, 3).

References

1. Bian L, Meng Y, Zhang M, Li D. (2019) MRE11-RAD50-NBS1 complex alterations and DNA damage response: implications for cancer treatment. Mol Cancer. 26;18(1):169. PMID: 31767017

2. Zhang Y, Zhou J, Lim CU. (2006) The role of NBS1 in DNA double strand break repair, telomere stability, and cell cycle checkpoint control. Cell Res. 16(1):45-54. PMID: 16467875

3. Komatsu K. NBS1 and multiple regulations of DNA damage response. (2016) J Radiat Res. 57 Suppl 1:i11-i17. PMID: 27068998

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 Nbs1 Antibody (NB100-143)(176)

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
(47)
IHC
(5)
IP
(18)
WB
(64)
All Applications
Filter By Species
Human
(94)
Mouse
(7)
Ape
(1)
Hamster
(1)
Mammal
(1)
Primate
(1)
All Species
Showing Publications 1 - 10 of 176. Show All 176 Publications.
Publications using NB100-143 Applications Species
Capper K. Targeting DNA Repair Mechanisms in MYC-Driven Cancer. Thesis. 2017 (WB, Mouse) WB Mouse
Howard SM, Ceppi I, Anand R et al. The internal region of CtIP negatively regulates DNA end resection Nucleic Acids Res. Apr 29 2020 [PMID: 32347940] (WB, Human) WB Human
Ragamin A, Yigit G, Bousset K et Al. Human RAD50 deficiency: Confirmation of a distinctive phenotype Am. J. Med. Genet. A Mar 25 2020 [PMID: 32212377] (WB, Human) WB Human
Qi F, Meng Q, Hayashi I, Kobayashi J FXR1 is a novel MRE11-binding partner and participates in oxidative stress responses J. Radiat. Res. Mar 25 2020 [PMID: 32211858] (WB, ICC/IF, IP, Human) WB, ICC/IF, IP Human
Mooser C, Symeonidou IE, Leimbacher PA et al. Treacle controls the nucleolar response to rDNA breaks via TOPBP1 recruitment and ATR activation Nat Commun Jan 8 2020 [PMID: 31913317] (ICC/IF, Human) ICC/IF Human
Volkening L, Vatselia A, Asgedom G et al. RAD50 regulates mitotic progression independent of DNA repair functions FASEB J. Dec 31 2019 [PMID: 31908056]
Yoshimura Y Increased error-free DNA repair gene expression through reprogramming in human iPS cells Regenerative Therapy 2019 Jun 28 [PMID: 31304203] (WB, Human) WB Human
Korsholm LM, Gal Z, Lin L et al. Double-strand breaks in ribosomal RNA genes activate a distinct signaling and chromatin response to facilitate nucleolar restructuring and repair Nucleic Acids Res. Jun 11 2019 [PMID: 31184714] (WB, ICC/IF, Human)

Details:
Induction of double strand breaks in human ribosomal RNA genes in the nucleolus is utilized to highlight the roles of the nucleolar DNA damage response.
WB, ICC/IF Human
Leimbacher PA, Jones SE, Shorrocks AK et al. MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis Mol. Cell Mar 18 2019 [PMID: 30898438] (WB, ICC/IF, Human) WB, ICC/IF Human
Qin B, Yu J, Nowsheen S et al. UFL1 promotes histone H4 ufmylation and ATM activation Nat Commun Mar 18 2019 [PMID: 30886146] (WB, Human) WB Human
Show All 176 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.

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
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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

Video Protocols

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

FAQs for Nbs1 Antibody (NB100-143). (Showing 1 - 8 of 8 FAQs).

  1. If this product is used in an application or species as a part of a customer review, will that validate this product in the application/species?
    • Yes, if this product is used in an untested application or species and it is shown to work through images from customer reviews or through publications, this validates the application/species for this product.
  2. What research areas can this product be used in?
    • All Nbs1 products can be used in Breast Cancer, Cancer, Checkpoint signaling, Chromatin Research, DNA Double Strand Break Repair, DNA Repair, Homologous Recombination, Non-homologous end-joining, Tumor Suppressors, and Cell Cycle and Replication.
  3. What is NBS1?
    • NBS1 is Nijmegen breakage syndrome protein 1, or Nibrin. Please refer to the background listed in the datasheet for a full summary of this common name.
  4. What is the theoretical molecular weight of Nbs1 antibodies?
    • In general, there is only one isoform described for this target. The TMW of the human form of the target protein is 85kDa. The mouse form is 84kDa. However, the observed molecular weight may vary.
  5. 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.
  6. 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.
  7. 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.
  8. I am doing ICC with Nbs1 antibody [NB100-143]. Do you have any recommendations on sample preparation?
    • Nbs1 protein is located in nuclei, so after fixation in 4% PFA, permeabilize the cells wurg 0.25% tritonX-100 in PBS, 10min room temperature. 

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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