Phospho-Rad17 (S656) Recombinant Rabbit Monoclonal Antibody [JE63-30]
Catalog# HA722148
Phospho-Rad17 (S656) Recombinant Rabbit Monoclonal Antibody [JE63-30]
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WB
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IF-Cell
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FC
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Human
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Mouse
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unconjugated
Overview
Product Name
Phospho-Rad17 (S656) Recombinant Rabbit Monoclonal Antibody [JE63-30]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide corresponding to residues surrounding Ser656 of human Rad17 protein.
Species Reactivity
Human, Mouse
Validated Applications
WB, IF-Cell, FC
Molecular Weight
Predicted band size: 77 kDa
Positive Control
HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, NIH/3T3 treated with 50ng/mL Calyculin A for 45 minutes cell lysate, HeLa cells treated with 100ng/mL Calyculin A for 30 minutes.
Conjugation
unconjugated
Clone Number
JE63-30
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IF-Cell
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1:500
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FC
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1:1,000
Target
Function
Cell cycle checkpoint protein RAD17 is a protein that in humans is encoded by the RAD17 gene. 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 ATR after the 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. Eight alternatively spliced transcript variants of this gene, which encode four distinct proteins, have been reported. Phospho-Rad17 (S656) Antibody [JE63-30] recognizes endogenous levels of Rad17 protein only when phosphorylated at Ser656.
Background References
1. Fukumoto Y et al. Rad17 Translocates to Nucleolus upon UV Irradiation through Nucleolar Localization Signal in the Central Basic Domain. Int J Mol Sci. 2022 Oct
2. Fukumoto Y et al. Nuclear translocation promotes proteasomal degradation of human Rad17 protein through the N-terminal destruction boxes. J Biol Chem. 2021 Aug
Subcellular Location
Nucleus.
Synonyms
CCYC antibody
Cell cycle checkpoint protein antibody
Cell cycle checkpoint protein RAD17 antibody
FLJ41520 antibody
HRAD 17 antibody
hRad17 antibody
R24L antibody
Rad 17 antibody
Rad 24 antibody
RAD1 (S. pombe) homolog antibody
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of Phospho-Rad17 (S656) on different lysates with Rabbit anti-Phospho-Rad17 (S656) antibody (HA722148) 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 λpp 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 λpp for 1 hour
Lysates/proteins at 20 µg/Lane.
Predicted band size: 77 kDa
Observed band size: 77 kDa
Exposure time: 43 seconds; ECL: K1802;
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 (HA722148) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of HeLa cells treated with or without 100ng/mL Calyculin A for 30 minutes labeling Phospho-Rad17 (S656) with Rabbit anti-Phospho-Rad17 (S656) antibody (HA722148) 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-Phospho-Rad17 (S656) antibody (HA722148) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. -
☑ Cell treatment (CT)
Flow cytometric analysis of HeLa cells treated with or without 100ng/mL Calyculin A for 30 minutes labeling Phospho-Rad17 (S656).
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722148, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"