Rad17 [p Ser656] Antibody (JE63-30)

Western Blot: Rad17 [p Ser656] Antibody (JE63-30) [NBP3-32792] - Western blot analysis of Rad17 on different lysates with Rabbit anti-Rad17 antibody (NBP3-32792) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate Lane 3: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, then the membrane treated with pp for 1 hour Lane 4: NIH/3T3 cell lysate Lane 5: NIH/3T3 treated with 50ng/mL Calyculin A for 45 minutes cell lysate Lane 6: NIH/3T3 treated with 50ng/mL Calyculin A for 45 minutes cell lysate, then the membrane treated with pp for 1 hour Lysates/proteins at 20 ug/Lane. Predicted band size: 77 kDa Observed band size: 77 kDa Exposure time: 43 seconds; ECL; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32792) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/ Immunofluorescence: Rad17 [p Ser656] Antibody (JE63-30) [NBP3-32792] - Immunocytochemistry analysis of HeLa cells treated with or without 100ng/mL Calyculin A for 30 minutes labeling Rad17 with Rabbit anti-Rad17 antibody (NBP3-32792) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Rad17 antibody (NBP3-32792) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Flow Cytometry: Rad17 [p Ser656] Antibody (JE63-30) [NBP3-32792] - Flow cytometric analysis of HeLa cells treated with or without 100ng/mL Calyculin A for 30 minutes labeling Rad17. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32792, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

• Reactivity: Hu, Mu
• Applications: WB, Flow, ICC/IF
• Clone: JE63-30
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Rad17 [p Ser656] Antibody (JE63-30) Summary

• Description: Novus Biologicals Rabbit Rad17 [p Ser656] Antibody (JE63-30) (NBP3-32792) is a recombinant monoclonal antibody validated for use in WB, Flow and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Synthetic peptide corresponding to residues surrounding Ser656 of human Rad17 protein. (Uniprot: O75943)
• Modification: p Ser656
• Localization: Nucleus.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: RAD17
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:500
  • Western Blot 1:1000
• Theoretical MW: 77 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for Rad17 [p Ser656] Antibody (JE63-30)

• 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

Background

Signals originating from damaged DNA activate checkpoint pathways and target the mitotic apparatus via a number a distinct mechanisms. These checkpoints link damage to DNA with the components of the cell cycle through signal transduction systems. In mammals, five genes have been identified as components of DNA damage-induced checkpoint pathways: ATM, p53, p21/WAF1/CIP1, hCHK1, and 14-3-3sigma. When overexpressed in mammalian cells, hRAD17 activates p53 and causes an accumulation of G1 phase cells, suggesting an involvement of the hRAD17 gene in this checkpoint pathway.(1).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: RAD17