Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human

Applications

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2 Alpha Clone # 7E4A2

Format

BSA Free
View all Nbs1 Primary Antibodies »

Product Specifications

Immunogen

Nbs1 Antibody (7E4A2) was made to partial recombinant human NBS1 (between residues 400-650) expressed in E. coli [Uniprot O60934]

Localization

Nucleus

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2 Alpha

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.

Scientific Data Images for Nbs1 Antibody (7E4A2) - BSA Free

Knockout Validated: Nbs1 Antibody (7E4A2) [NBP2-26297]

Western Blot: Nbs1 Antibody (7E4A2) [NBP2-26297]

Western Blot: Nbs1 Antibody (7E4A2) [NBP2-26297] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Nbs1 knockout (KO) HeLa cell line. PVDF membrane was probed with 1:1000 of Mouse Anti-Human Polyclonal Nbs1 Antibody [NBP2-26297] followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody [HAF018]. Specific band was detected for Nbs1 at approximately 95 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.
Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody (7E4A2) [NBP2-26297]

Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody (7E4A2) [NBP2-26297]

Immunocytochemistry/Immunofluorescence: Nbs1 Antibody (7E4A2) [NBP2-26297] - Analysis of Hela cells using mouse Nbs1 Antibody (7E4A2) [NBP2-26297] (green). Blue: DRAQ5 fluorescent DNA dye [NBP2-81125]. Red: Actin filaments have been labeled with Alexa Fluor-555 phalloidin.
Immunohistochemistry: Nbs1 Antibody (7E4A2) [NBP2-26297]

Immunohistochemistry: Nbs1 Antibody (7E4A2) [NBP2-26297]

Immunohistochemistry: Nbs1 Antibody (7E4A2) [NBP2-26297] - Analysis of paraffin-embedded rectum cancer tissues using mouse Nbs1 Antibody (7E4A2) [NBP2-26297] with DAB staining.
Flow Cytometry: Nbs1 Antibody (7E4A2) [NBP2-26297]

Flow Cytometry: Nbs1 Antibody (7E4A2) [NBP2-26297]

Flow Cytometry: Nbs1 Antibody (7E4A2) [NBP2-26297] - Analysis of Nbs1 in Hela cells using mouse Nbs1 Antibody (7E4A2) [NBP2-26297] (green) and negative control (red).
Immunohistochemistry-Paraffin: Nbs1 Antibody (7E4A2) [NBP2-26297]

Immunohistochemistry-Paraffin: Nbs1 Antibody (7E4A2) [NBP2-26297]

Immunohistochemistry-Paraffin: Nbs1 Antibody (7E4A2) [NBP2-26297] - Analysis of Nbs1 in paraffin-embedded cervical cancer tissues using mouse Nbs1 Antibody (7E4A2) [NBP2-26297] with DAB staining.
ELISA: Nbs1 Antibody (7E4A2) [NBP2-26297]

ELISA: Nbs1 Antibody (7E4A2) [NBP2-26297]

ELISA: Nbs1 Antibody (7E4A2) [NBP2-26297] - ELISA analysis using Nbs1 Antibody (7E4A2) [NBP2-26297].
Simple Western: Nbs1 Antibody (7E4A2) [NBP2-26297]

Simple Western: Nbs1 Antibody (7E4A2) [NBP2-26297]

Simple Western: Nbs1 Antibody (7E4A2) [NBP2-26297] - Analysis using Nbs1 Antibody (7E4A2) [NBP2-26297]. Molecular Weight observed at: 115 kDa.

Applications for Nbs1 Antibody (7E4A2) - BSA Free

Application
Recommended Usage

Flow Cytometry

1:200 - 1:400

Immunocytochemistry/ Immunofluorescence

1-50 - 1:100

Immunohistochemistry

1:200 - 1:1000

Simple Western

1:50

Western Blot

1:500 - 1:2000
Application Notes
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HepG2 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:50, apparent MW was 71 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Ammonium sulfate precipitation

Formulation

PBS, 0.5% proprietary protein stabilizer

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 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

Entrez Gene IDs

4683 (Human)

Gene Symbol

NBN

UniProt

O60934

Additional Nbs1 Products

Product Documents for Nbs1 Antibody (7E4A2) - BSA Free

Certificate of Analysis

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

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Product Specific Notices for Nbs1 Antibody (7E4A2) - 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

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Protocols

View specific protocols for Nbs1 Antibody (7E4A2) - BSA Free (NBP2-26297):


Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute anti-NBS1 primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted 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 three 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.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

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

FAQs for Nbs1 Antibody (7E4A2) - BSA Free

Showing  1 - 5 of 5 FAQs Showing All

  • Q: How was this antibody purified?

    A: Nbs1 Antibody (7E4A2) was purified by ammonium sulfate precipitation.

  • 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: How was this antibody purified?

    A: Nbs1 Antibody (7E4A2) was purified by ammonium sulfate precipitation.

  • 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: How was this antibody purified?

    A: Nbs1 Antibody (7E4A2) was purified by ammonium sulfate precipitation.

  • 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: How was this antibody purified?

    A: Nbs1 Antibody (7E4A2) was purified by ammonium sulfate precipitation.

  • 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: How was this antibody purified?

    A: Nbs1 Antibody (7E4A2) was purified by ammonium sulfate precipitation.

  • 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 - 5 of 5 FAQs Showing All
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