Nbs1 Overexpression Lysate

Novus Biologicals | Catalog # NBL1-13497

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Nbs1 Overexpression Lysate

Nbs1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC214682

Accession#: NM_002485

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Theoretical MW

84.8 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.

Type

Overexpression

Scientific Data Images for Nbs1 Overexpression Lysate

Western Blot: Nbs1 Overexpression Lysate [NBL1-13497]

Western Blot: Nbs1 Overexpression Lysate [NBL1-13497]

Western Blot: Nbs1 Overexpression Lysate [NBL1-13497] - Western blot validation of Nbs1 Overexpression Lysate [NBL1-13497] using anti-DDK antibody [NBP1-71705]. Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with p95 Nbs1 (NBN) (NM_002485) Human Tagged ORF Clone using transfection reagent MegaTran 2.0. Observed molecular weight at ~95 kDa.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. 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 Overexpression Lysate

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 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

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.

Related Research Areas

DNA Repair

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FAQs for Nbs1 Overexpression Lysate

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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