AB220217

Anti-p95/NBS1 antibody [Y112] - BSA and Azide free

RabMAb
Recombinant
KO Validated
What is this?

Be the first to review this product! Submit a review

(1 Publication)

Rabbit Recombinant Monoclonal p95/NBS1 antibody. Carrier free. Suitable for IHC-P, WB and reacts with Human samples. Cited in 1 publication.

4 Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling p95/NBS1 with purified ab32074 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32074).

IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling p95/NBS1 with unpurified ab32074 at 1/20. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32074).

Lab

Western blot - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

This data was developed using the same antibody clone in a different buffer formulation (ab32074).

Lanes 1 - 2 : Merged signal (red and green). Green - ab32074 observed at 90 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab32074 was shown to react with p95/NBS1 in wild-type A431 cells in Western blot with loss of signal observed in NBN knockout cell line ab269506 (NBN knockout cell lysate ab269668). Wild-type A431 and NBN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween^®) before incubation with ab32074 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

Lanes 1 - 2:

Western blot - Anti-p95/NBS1 antibody [Y112] (<a href='/en-us/products/primary-antibodies/p95-nbs1-antibody-y112-ab32074'>ab32074</a>) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-p95/NBS1 antibody (<a href='/en-us/products/primary-antibodies/p95-nbs1-antibody-ab23996'>ab23996</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

Western blot - Human NBN (p95/NBS1) knockout A-431 cell lysate (<a href='/en-us/products/cell-lysates/human-nbn-p95-nbs1-knockout-a-431-cell-lysate-ab269668'>ab269668</a>) at 20 µg

Predicted band size: 84 kDa

Observed band size: 90 kDa,95 kDa

false

Lab

Western blot - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

This data was developed using the same antibody clone in a different buffer formulation (ab32074).

Lanes 1-3 : Merged signal (red and green). Green - ab32074 observed at 95 kDa.

ab32074 Anti-p95/NBS1 antibody [Y112] was shown to specifically react with p95/NBS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261834 (knockout cell lysate ab257111) was used. Wild-type and p95/NBS1 knockout samples were subjected to SDS-PAGE. ab32074, Anti-GAPDH antibody [6C5] - Cytoplasmic Loading Control (ab8245) and Anti-Histone H3 (ab176842) - Nuclear Loading Control were incubated overnight at 4°C at 1 in 1000 dilution, 1 in 20000 dilution and 1 in 1000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773), Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-p95/NBS1 antibody [Y112] (<a href='/en-us/products/primary-antibodies/p95-nbs1-antibody-y112-ab32074'>ab32074</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

NBN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human NBN (p95/NBS1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-nbn-p95-nbs1-knockout-hela-cell-line-ab261834'>ab261834</a>)

Lane 3:

Wild-type HeLa nuclear cell lysate at 20 µg

Predicted band size: 84 kDa

Observed band size: 95 kDa

false

Unconjugated

Anti-p95/NBS1 antibody [Y112]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

Y112

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

IHC-P, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

style

left-column

style

left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-low-endotoxin-azide-free-ab199376'>ab199376</a> - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Mouse": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Rat": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }

style

left-column

Product details

ab220217 is the carrier-free version of ab32074.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

style

left-column

Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

style

left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The target known as p95/NBS1 also referred to as NBN protein is a critical component in the DNA damage response mechanism. The full molecular weight of the NBS1 protein is approximately 95 kDa. This protein is abundantly expressed in various tissues particularly where there is a higher rate of cell division or repair such as in the thymus and testis. It plays an important role in maintaining the stability of the genome through its involvement in the repair of double-strand breaks.

Biological function summary

The NBS1 protein functions as an essential component of the MRN complex which also includes MRE11 and RAD50 proteins. This complex is fundamental in accurately detecting DNA double-strand breaks and initiating repair processes. Through its actions NBS1 facilitates numerous cellular processes that preserve genomic integrity such as homologous recombination and non-homologous end joining. Its influence in controlling the cell cycle further emphasizes its role in maintaining cellular health.

Pathways

The NBS1 protein engages significantly in the DNA damage response and repair pathways notably impacting the ATM signaling pathway. By interacting with proteins such as ATM kinase NBS1 facilitates the phosphorylation and activation of several downstream effectors necessary for DNA repair and cell cycle checkpoints. The MRN complex also links with the RAD50 protein highlighting its involvement in these pathways.

Mutations in the NBS1 gene are linked to Nijmegen breakage syndrome characterized by sensitivity to radiation immune deficiency and increased cancer risk. Additionally NBS1's role in pathways makes it pertinent to certain cancer pathologies where its malfunction can contribute to genomic instability. The NBN protein through its interaction with ATM and RAD50 assists in understanding its connection with predispositions to these diseases.

style

left-column

style

left-column

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

style

left-column

Target data

Component of the MRN complex, which plays a central role in double-strand break (DSB) repair, DNA recombination, maintenance of telomere integrity and meiosis (PubMed : 10888888, PubMed : 15616588, PubMed : 18411307, PubMed : 18583988, PubMed : 18678890, PubMed : 19759395, PubMed : 23115235, PubMed : 28216226, PubMed : 28867292, PubMed : 9705271). The MRN complex is involved in the repair of DNA double-strand breaks (DSBs) via homologous recombination (HR), an error-free mechanism which primarily occurs during S and G2 phases (PubMed : 19759395, PubMed : 28867292, PubMed : 9705271). The complex (1) mediates the end resection of damaged DNA, which generates proper single-stranded DNA, a key initial steps in HR, and is (2) required for the recruitment of other repair factors and efficient activation of ATM and ATR upon DNA damage (PubMed : 19759395, PubMed : 9705271). The MRN complex possesses single-strand endonuclease activity and double-strand-specific 3'-5' exonuclease activity, which are provided by MRE11, to initiate end resection, which is required for single-strand invasion and recombination (PubMed : 19759395, PubMed : 28867292, PubMed : 9705271). Within the MRN complex, NBN acts as a protein-protein adapter, which specifically recognizes and binds phosphorylated proteins, promoting their recruitment to DNA damage sites (PubMed : 12419185, PubMed : 15616588, PubMed : 18411307, PubMed : 18582474, PubMed : 18583988, PubMed : 18678890, PubMed : 19759395, PubMed : 19804756, PubMed : 23762398, PubMed : 24534091, PubMed : 27814491, PubMed : 27889449, PubMed : 33836577). Recruits MRE11 and RAD50 components of the MRN complex to DSBs in response to DNA damage (PubMed : 12419185, PubMed : 18411307, PubMed : 18583988, PubMed : 18678890, PubMed : 24534091, PubMed : 26438602). Promotes the recruitment of PI3/PI4-kinase family members ATM, ATR, and probably DNA-PKcs to the DNA damage sites, activating their functions (PubMed : 15064416, PubMed : 15616588, PubMed : 15790808, PubMed : 16622404, PubMed : 22464731, PubMed : 30952868, PubMed : 35076389). Mediates the recruitment of phosphorylated RBBP8/CtIP to DSBs, leading to cooperation between the MRN complex and RBBP8/CtIP to initiate end resection (PubMed : 19759395, PubMed : 27814491, PubMed : 27889449, PubMed : 33836577). RBBP8/CtIP specifically promotes the endonuclease activity of the MRN complex to clear DNA ends containing protein adducts (PubMed : 27814491, PubMed : 27889449, PubMed : 30787182, PubMed : 33836577). The MRN complex is also required for the processing of R-loops (PubMed : 31537797). NBN also functions in telomere length maintenance via its interaction with TERF2 : interaction with TERF2 during G1 phase preventing recruitment of DCLRE1B/Apollo to telomeres (PubMed : 10888888, PubMed : 28216226). NBN also promotes DNA repair choice at dysfunctional telomeres : NBN phosphorylation by CDK2 promotes non-homologous end joining repair at telomeres, while unphosphorylated NBN promotes microhomology-mediated end-joining (MMEJ) repair (PubMed : 28216226). Enhances AKT1 phosphorylation possibly by association with the mTORC2 complex (PubMed : 23762398).

See full target information NBN

Alternative Names

NBS, NBS1, P95, NBN, Nibrin, Cell cycle regulatory protein p95, Nijmegen breakage syndrome protein 1, hNbs1

style

left-column

Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Brain : a journal of neurology 137:3235-47 PubMed25351739

2014

Augmentation of phenotype in a transgenic Parkinson mouse heterozygous for a Gaucher mutation.

Applications

Unspecified application

Species

Unspecified reactive species

Ianai Fishbein,Yien-Ming Kuo,Benoit I Giasson,Robert L Nussbaum

View all publications

style

left-column

style

left-column

style

right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-p95/NBS1 antibody [Y112] - BSA and Azide free

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

Western blot - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

Western blot - Anti-p95/NBS1 antibody [Y112] - BSA and Azide free (AB220217)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Alternative Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Alternative Names

Publications (1)

Product promise