Anti-Rad17 Antibody Picoband™

Product Name

Anti-Rad17 Antibody Picoband™

View all RAD17 Antibodies

SKU/Catalog Number

A02159-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Rad17 Antibody Picoband™ catalog # A02159-2. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

Storage & Handling

Store 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 freeze-thaw cycles.

Cite This Product

Anti-Rad17 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02159-2)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse Rad17 recombinant protein (Position: M1-Q267).

*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

A02159-2 is reactive to RAD17 in Mouse, Rat

Applications

A02159-2 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

77 kDa

Calculated molecular weight

77.055kDa

Background of RAD17

The protein encoded by this gene is highly similar to the gene product of Schizosaccharomyces pombe rad17, a cell cycle checkpoint gene required for cell cycle arrest and DNA damage repair in response to DNA damage. This protein shares strong similarity with DNA replication factor C (RFC), and can form a complex with RFCs. This protein binds to chromatin prior to DNA damage and is phosphorylated by the checkpoint kinase ATR following damage. This protein recruits the RAD1-RAD9-HUS1 checkpoint protein complex onto chromatin after DNA damage, which may be required for its phosphorylation. The phosphorylation of this protein is required for the DNA-damage-induced cell cycle G2 arrest, and is thought to be a critical early event during checkpoint signaling in DNA-damaged cells. Multiple alternatively spliced transcript variants of this gene, which encode four distinct protein isoforms, have been reported. Two pseudogenes, located on chromosomes 7 and 13, have been identified.

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

Add 0.2ml of distilled water will yield a concentration of 500ug/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, 0.25-0.5μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat
Flow Cytometry, 1-3μg/1x10⁶ cells, Mouse
Direct ELISA, 0.1-0.5μg/ml, Mouse

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Rad17 using anti-Rad17 antibody (A02159-2).
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 50ug of sample under reducing conditions.
Lane 1: mouse heart tissue lysates,
Lane 2: mouse NIH/3T3 whole cell lysates,
Lane 3: rat heart tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Rad17 antigen affinity purified polyclonal antibody (Catalog # A02159-2) 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 1:10000 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 Rad17 at approximately 77KD. The expected band size for Rad17 is at 77KD.

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Figure 2. IHC analysis of Rad17 using anti-Rad17 antibody (A02159-2).
Rad17 was detected in paraffin-embedded section of mouse Peyer's patches tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Rad17 Antibody (A02159-2) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of Rad17 using anti-Rad17 antibody (A02159-2).
Rad17 was detected in paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Rad17 Antibody (A02159-2) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of HEPA1-6 cells using anti-Rad17 antibody (A02159-2).
Overlay histogram showing HEPA1-6 cells stained with A02159-2 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Rad17 Antibody (A02159-2,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 5. Flow Cytometry analysis of mouse spleen tissues using anti-Rad17 antibody (A02159-2).
Overlay histogram showing mouse spleen tissues stained with A02159-2 (Blue line).The tissues were blocked with 10% normal goat serum. And then incubated with rabbit anti-Rad17 Antibody (A02159-2,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For RAD17 (Source: Uniprot.org, NCBI)

Alternative Names

CCYC; cell cycle checkpoint protein (RAD17); cell cycle checkpoint protein RAD17; FLJ41520; HRAD17; R24L; RAD1 (S. pombe) homolog; RAD1 homolog; RAD17 homolog (S. pombe); Rad17; Rad17-like protein; RAD17SP; RAD24; RF-C activator 1 homolog; RF-C/activator 1 homolog RAD17 CCYC, HRAD17, R24LSP, RAD24, RAD17 RAD17 checkpoint clamp loader component cell cycle checkpoint protein RAD17|RAD1 homolog|RAD17 homolog|RF-C activator 1 homolog|Rad17-like protein|cell cycle checkpoint protein (RAD17)

For more info on RAD17, check out the RAD17 Infographic

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In this infographic, you will see the following information for RAD17: 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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