Nbs1 Knockout HeLa Cell Lysate

Product Discontinued

Nbs1 Knockout HeLa Cell Lysate Summary

• Preparation Method: Knockout achieved by using CRISPR/Cas9,-16 bp deletion in exon2 and 1 bp insertion in exon2
• Gene: NBN

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines.
• Concentration: LYOPH
• Reconstitution Instructions: To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel.

Lysate Details for Array

• Type: Knockout HeLa Cell

Notes

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Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.

Alternate Names for Nbs1 Knockout HeLa Cell Lysate

• 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. Lysates are guaranteed for 6 months from date of receipt.

FAQs for Nbs1 Lysate (NBP2-65446). (Showing 1 - 4 of 4 FAQ).

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.

Bioinformatics

• Gene Symbol: NBN