Anti-p95 NBS1 Antibody Picoband™ (monoclonal, 5D7A1)

Product Name

Anti-p95 NBS1 Antibody Picoband™ (monoclonal, 5D7A1)

View all Nbs1 Antibodies

SKU/Catalog Number

M00732-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-p95 NBS1 Antibody Picoband™ (monoclonal, 5D7A1) catalog # M00732-1. Tested in FCM, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-p95 NBS1 Antibody Picoband™ (monoclonal, 5D7A1) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00732-1)

Host

Mouse

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Clonality

Monoclonal

Clone Number

5D7A1

Isotype

Mouse IgG2a

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of human p95 NBS1, different from the related mouse sequence by three amino acids, and from the related rat sequence by five amino acids.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M00732-1 is reactive to NBN in Human, Mouse, Rat

Applications

M00732-1 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

95 kDa

Calculated molecular weight

84.959kDa

Background of Nbs1

p95 NBS1, also known as NBN or Nibrin, is a protein which in humans is encoded by the NBN gene. Nibrin is a protein associated with the repair of double strand breaks (DSBs) which pose serious damage to a genome. It is a 754 amino acid protein identified as a member of the NBS1/hMre11/RAD50(N/M/R, more commonly referred to asMRN) double strand DNA break repair complex. This complex recognizes DNA damage and rapidly relocates to DSB sites and forms nuclear foci. It also has a role in regulation of N/M/R (MRN) protein complex activity which includes end-processing of both physiological and mutagenic DNA double strand breaks (DSBs).

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of p95 NBS1 using anti-p95 NBS1 antibody (M00732-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: rat brain tissue lysates,
Lane 4: rat testis tissue lysates,
Lane 5: mouse brain tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-p95 NBS1 antigen affinity purified monoclonal antibody (Catalog # M00732-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for p95 NBS1 at approximately 95 kDa. The expected band size for p95 NBS1 is at 95 kDa.

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Figure 2. IHC analysis of p95 NBS1 using anti-p95 NBS1 antibody (M00732-1).
p95 NBS1 was detected in a paraffin-embedded section of human testis cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml mouse anti-p95 NBS1 Antibody (M00732-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

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Figure 3. IF analysis of p95 NBS1 using anti-p95 NBS1 antibody (M00732-1).
p95 NBS1 was detected in an immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL mouse anti-p95 NBS1 Antibody (M00732-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 4. Flow Cytometry analysis of A431 cells using anti-p95 NBS1 antibody (M00732-1).
Overlay histogram showing A431 cells stained with M00732-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-p95 NBS1 Antibody (M00732-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For NBN (Source: Uniprot.org, NCBI)

Alternative Names

ATV; AT-V1; AT-V2; Cell cycle regulatory protein p95; FLJ10155; MGC87362; NBN; Nbs1; NBS1P95; NBSNijmegen breakage syndrome protein 1; Nibrin; Nijmegen breakage syndrome 1 (nibrin); p95 protein of the MRE11/RAD50 complex; p95 NBN AT-V1, AT-V2, ATV, NBS, NBS1, P95 nibrin nibrin|Nijmegen breakage syndrome 1 (nibrin)|cell cycle regulatory protein p95|p95 protein of the MRE11/RAD50 complex

For more info on NBN, check out the NBN Infographic

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In this infographic, you will see the following information for NBN: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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