Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Cited:

Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Nbs1 Primary Antibodies »

Product Specifications

Immunogen

Nbs1 Antibody was made to a synthetic peptide made to an internal portion of the mouse NBS1 protein (within residues 150-200). [Swiss-Prot# Q9R207]

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 21700704).

Localization

Nucleus. Telomere. Note= Localizes to discrete nuclear foci after treatment with genotoxic agents.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

84 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.

Scientific Data Images for Nbs1 Antibody - BSA Free

Western Blot: Nbs1 AntibodyBSA Free [NBP1-06705]

Western Blot: Nbs1 AntibodyBSA Free [NBP1-06705]

Western Blot: Nbs1 Antibody [NBP1-06705] - Western Blot analysis of Nbs1 on NIH/3T3 (Lane 6) and HeLa whole cell extract (Lane 7) with Nbs1 Antibody [NBP1-06705]. Observed molecular weight at ~99 kDa.
Immunohistochemistry-Paraffin: Nbs1 Antibody - BSA Free [NBP1-06705]

Immunohistochemistry-Paraffin: Nbs1 Antibody - BSA Free [NBP1-06705]

Immunohistochemistry-Paraffin: Nbs1 Antibody [NBP1-06705] - Staining of paraffin-embedded mouse skin with Nbs1 Antibody [NBP1-06705].

Applications for Nbs1 Antibody - BSA Free

Application
Recommended Usage

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Western Blot

0.5 ug/ml. Use reported in scientific literature (PMID 21700704)
Application Notes
In Western blot, a band is seen at ~97 kDa.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.1% Sodium Azide

Concentration

1.48 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Nbs1

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

Long Name

Nijmegen Breakage Syndrome 1

Alternate Names

NBN, Nibrin, p95

Gene Symbol

NBN

Additional Nbs1 Products

Product Documents for Nbs1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Nbs1 Antibody - BSA Free

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.

Related Research Areas

DNA Repair

Citations for Nbs1 Antibody - BSA Free

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Protocols

View specific protocols for Nbs1 Antibody - BSA Free (NBP1-06705):

Nbs1 Antibody:
Immunohistochemistry-paraffin embedded sections

Antigen Unmasking
Bring slides to a boil in 10 mM sodium citrate buffer pH 6.0 then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench top for 30 minutes.

Staining
1. Wash sections in dH2O three times for 5 minutes each.
2. Wash section in wash buffer (1X PBS/0.1% Tween-20 (1X PBST)) for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1X PBST, 5% goat serum) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul primary antibody diluted in 1X PBST, 5% goat serum to each section. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated secondary antibody, diluted in 1X PBST, 5% goat serum. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Striptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in dH2O.
12. Counterstain sections in hematoxylin.
13. Wash sections in dH2O two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

Nbs1 Antibody:
Procedure Guide for NPB1-06705 - NBS1 Antibody
Western Blot Protocol

1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 30 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% NFDM + 1% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-NBS1 primary antibody (NBP1-06705) in blocking buffer and incubate 1 hour at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).

**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

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

FAQs for Nbs1 Antibody - BSA Free

Showing  1 - 4 of 4 FAQs Showing All

  • Q: 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?

    A: 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.

  • Q: What is NBS1?

    A: 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.

  • Q: What is the theoretical molecular weight of Nbs1 antibodies?

    A: 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.

  • Q: What research areas can this product be used in?

    A: 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.

  • Q: 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?

    A: 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.

  • Q: What is NBS1?

    A: 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.

  • Q: What is the theoretical molecular weight of Nbs1 antibodies?

    A: 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.

  • Q: What research areas can this product be used in?

    A: 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.

  • Q: 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?

    A: 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.

  • Q: What is NBS1?

    A: 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.

  • Q: What is the theoretical molecular weight of Nbs1 antibodies?

    A: 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.

  • Q: What research areas can this product be used in?

    A: 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.

  • Q: 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?

    A: 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.

  • Q: What is NBS1?

    A: 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.

  • Q: What is the theoretical molecular weight of Nbs1 antibodies?

    A: 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.

  • Q: What research areas can this product be used in?

    A: 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.

Showing  1 - 4 of 4 FAQs Showing All
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