Rabbit anti-Mouse, Rat Rad17 Polyclonal Antibody | anti-Rad17 antibody

Anti-Rad17 Antibody

Cat. Num. AAA125638

• Gene Names: Rad17; MmRad24
• Reactivity: Mouse, Rat
• Applications: ELISA, Flow Cytometry, Functional Assay, Immunohistochemistry, Western Blot
• Purity: Immunogen affinity purified.
• Synonyms: Rad17, Antibody; Anti-Rad17 Antibody; Cell cycle checkpoint protein RAD17; Rad17; RAD17 checkpoint clamp loader component; anti-Rad17 antibody

• Host: Rabbit
• Reactivity: Mouse, Rat
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Rabbit IgG polyclonal antibody for Rad17 detection.
• Purity/Purification: Immunogen affinity purified.
• Form/Format: Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.

• Applicable Applications for anti-Rad17 antibody: ELISA (Direct ELISA), FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot)
• Immunogen: E Coli-derived mouse Rad17 recombinant protein (Position: M1-Q267).
• Reconstitution: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Recommended Detection Systems: Recommended Detection Systems
• Preparation and Storage: Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

FCM/FACS (Flow Cytometry)

(Figure 5. Flow Cytometry analysis of mouse spleen tissues using anti-Rad17 antibody (AAA125638).Overlay histogram showing mouse spleen tissues stained with AAA125638 (Blue line). The tissues were blocked with 10% normal goat serum. And then incubated with rabbit anti-Rad17 Antibody (AAA125638,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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.)

FCM/FACS (Flow Cytometry)

(Figure 4. Flow Cytometry analysis of HEPA1-6 cells using anti-Rad17 antibody (AAA125638).Overlay histogram showing HEPA1-6 cells stained with AAA125638 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Rad17 Antibody (AAA125638,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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.)

IHC (Immunohistochemisry)

(Figure 3. IHC analysis of Rad17 using anti-Rad17 antibody (AAA125638).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 (AAA125638) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohiostchemistry)

(Figure 2. IHC analysis of Rad17 using anti-Rad17 antibody (AAA125638).Rad17 was detected in paraffin-embedded section of mouse Peyers 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 (AAA125638) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of Rad17 using anti-Rad17 antibody (AAA125638).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 lysatesLane 2: mouse NIH/3T3 whole cell lysatesLane 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 # AAA125638) at 0. 5 μg/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Rad17 at approximately 77KD. The expected band size for Rad17 is at 77KD.)

Related Product Information for anti-Rad17 antibody

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.

References

1. Bao, S., Shen, X., Shen, K., Liu, Y., Wang, X. -F. The mammalian Rad24 homologous to yeast Saccharomyces cerevisiae Rad24 and Schizosaccharomyces pombe Rad17 is involved in DNA damage checkpoint. Cell Growth Diff. 9: 961-967, 1998.
2. Bao, S., Tibbetts, R. S., Brumbaugh, K. M., Fang, Y., Richardson, D. A., Ali, A., Chen, S. M., Abraham, R. T., Wang, X. -F. ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses. Nature 411: 969-974, 2001.
3. Bluyssen, H. A. R., Naus, N. C., van Os, R. I., Jaspers, I., Hoeijmakers, J. H. J., de Klein, A. Human and mouse homologues of the Schizosaccharomyces pombe rad17+ cell cycle checkpoint control gene. Genomics 55: 219-228, 1999.

NCBI and Uniprot Product Information

• NCBI GI #: 113199779
• NCBI GeneID: 19356
• NCBI Accession #: NP_001037836.1
• NCBI GenBank Nucleotide #: NM_001044371.2
• UniProt Accession #: Q6NXW6; O88934; O89024
• Molecular Weight: 77,391 Da
• NCBI Official Full Name: cell cycle checkpoint protein RAD17 isoform 1
• NCBI Official Synonym Full Names: RAD17 homolog (S. pombe)
• NCBI Official Symbol: Rad17
• NCBI Official Synonym Symbols: MmRad24
• NCBI Protein Information: cell cycle checkpoint protein RAD17
• UniProt Protein Name: Cell cycle checkpoint protein RAD17
• UniProt Gene Name: Rad17
• UniProt Entry Name: RAD17_MOUSE