Anti-RAD50 Antibody Picoband™

Product Name

Anti-RAD50 Antibody Picoband™

View all RAD50 Antibodies

SKU/Catalog Number

A00347-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-RAD50 Antibody Picoband™ catalog # A00347-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

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-RAD50 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00347-2)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human RAD50 recombinant protein (Position: N1224-H1312).

*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

A00347-2 is reactive to RAD50 in Human

Applications

A00347-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

154 kDa

Calculated molecular weight

153.892kDa

Background of RAD50

DNA repair protein RAD50, also known as RAD50, is a protein that in humans is encoded by the RAD50 gene. The protein encoded by this gene is highly similar to Saccharomyces cerevisiae Rad50, a protein involved in DNA double-strand break repair. This protein forms a complex with MRE11 and NBS1. The protein complex binds to DNA and displays numerous enzymatic activities that are required for nonhomologous joining of DNA ends. This protein, cooperating with its partners, is important for DNA double-strand break repair, cell cycle checkpoint activation, telomere maintenance, and meiotic recombination. Knockout studies of the mouse homolog suggest this gene is essential for cell growth and viability. Mutations in this gene are the cause of Nijmegen breakage syndrome-like disorder.

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
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of RAD50 using anti-RAD50 antibody (A00347-2).
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 MCF-7 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: human A549 whole cell lysates,
Lane 6: human Caco-2 whole cell lysates,
Lane 7: human Daudi whole cell lysates,
Lane 8: human SH-SY5Y whole cell 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 rabbit anti-RAD50 antigen affinity purified polyclonal antibody (Catalog # A00347-2) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for RAD50 at approximately 154 kDa. The expected band size for RAD50 is at 154 kDa.

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Figure 2. Flow Cytometry analysis of A549 cells using anti-RAD50 antibody (A00347-2).
Overlay histogram showing A549 cells stained with A00347-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-RAD50 Antibody (A00347-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 3. IF analysis of RAD50 using anti-RAD50 antibody (A00347-2).
RAD50 was detected in an immunocytochemical section of A549 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 rabbit anti-RAD50 Antibody (A00347-2) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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.

Gene/Protein Information For RAD50 (Source: Uniprot.org, NCBI)

Alternative Names

DNA repair protein RAD50; EC 3.6; EC 3.6.1.15; EC 3.6.3; EC 3.6.3.27; hRad50; NBSLD; RAD50 (S. cerevisiae) homolog; RAD50 homolog (S. cerevisiae); Rad50; RAD502; RAD50-2 RAD50 NBSLD2, hRad50, RAD50 RAD50 double strand break repair protein DNA repair protein RAD50|RAD50 homolog, double strand break repair protein

For more info on RAD50, check out the RAD50 Infographic

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In this infographic, you will see the following information for RAD50: 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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