Anti-p95/NBS1 antibody [Y112] (ab32074)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y112] to p95/NBS1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-p95/NBS1 antibody [Y112]
See all p95/NBS1 primary antibodies -
Description
Rabbit monoclonal [Y112] to p95/NBS1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human -
Immunogen
Synthetic peptide within Human p95/NBS1 aa 650-750 (C terminal). The exact sequence is proprietary.
Database link: O60934 -
Positive control
- WB: HeLa and Jurkat cell lysates. IHC-P: Human testis and skin carcinoma tissues.
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General notes
This product was previously labelled as p95 NBS1
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y112 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-p95/NBS1 antibody [Y112] (ab32074) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NBN knockout HeLa cell lysate
Lane 3 : Wild-type HeLa nuclear cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?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.
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All lanes : Anti-p95/NBS1 antibody [Y112] (ab32074) at 1/1200 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 85 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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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.
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All lanes : Anti-p95/NBS1 antibody [Y112] (ab32074) at 1/10000 dilution (purified)
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 85 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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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.
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