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Rad17 [p Ser656] Antibody (JE63-30)

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32792

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne
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NBP3-32792

Key Product Details

Species Reactivity

Validated:

Human, Mouse

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # JE63-30

Concentration

1 mg/ml

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

Immunogen

Synthetic peptide corresponding to residues surrounding Ser656 of human Rad17 protein. (Uniprot: O75943)

Modification

p Ser 656

Localization

Nucleus.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

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.

Scientific Data Images for Rad17 [p Ser656] Antibody (JE63-30)

Rad17 [p Ser656] Antibody (JE63-30)

Western Blot: Rad17 [p Ser656] Antibody (JE63-30) [NBP3-32792]

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

Immunocytochemistry/ Immunofluorescence: Rad17 [p Ser656] Antibody (JE63-30) [NBP3-32792]

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

Flow Cytometry: Rad17 [p Ser656] Antibody (JE63-30) [NBP3-32792]

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).

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

Application
Recommended Usage

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

1:500

Western Blot

1:1000
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

1*TBS (pH7.4), 0.05% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Rad17

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).

Alternate 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-like protein, RAD17Sp, Rad24, RF-C activator 1 homolog, RF-C/activator 1 homolog

Gene Symbol

RAD17

Additional Rad17 Products

Product Documents for Rad17 [p Ser656] Antibody (JE63-30)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Rad17 [p Ser656] Antibody (JE63-30)

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.

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