RAD17 antibody - N-terminal region (ARP30546_P050)

Catalog#: ARP30546_P050

• Size: 50ug

Human Fetal liver

Human Ovcar-3

• Description of Target: RAD17 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.
• Gene Symbol: RAD17
• Official Gene Full Name: RAD17 homolog (S. pombe)
• Alias Symbols: CCYC; FLJ41520; HRAD17; R24L; RAD17SP; RAD24
• Protein Accession#: NP_002864
• Nucleotide Accession#: NM_002873
• Swissprot Id: O75943-2
• Host: Rabbit
• Clonality: Polyclonal
• Protein Size (# AA): 670
• Molecular Weight: 76kDa
• Application: WB
• Partner Proteins: ATM,ATR,HUS1,MCM7,NHP2L1,POLE,RAD1,RAD9A,RAD9B,RFC4,ALK,ATM,ATR,CLSPN,FOXO3,H2AFX,HUS1,NHP2L1,POLE,POLE2,POLE3,POLE4,PRKDC,RAD1,RAD9A,RAD9B,RFC1,RFC2,RFC3,RFC4,RFC5,USP20
• Immunogen: The immunogen for anti-RAD17 antibody: synthetic peptide directed towards the N terminal of human RAD17
• Product Format: Lyophilized powder
• Purification: Affinity Purified
• Predicted Homology Based on Immunogen Sequence: Dog: 100%; Human: 100%; Pig: 100%; Horse: 91%
• Peptide Sequence: Synthetic peptide located within the following region: MNQVTDWVDPSFDDFLECSGVSTITATSLGVNNSSHRRKNGPSTLESSRF
• Blocking Peptide: For anti-RAD17 antibody is Catalog # AAP30546 (Previous Catalog # AAPP01193)
• Key Reference: N/A
• Reconstitution and Storage: Add 50 μl of distilled water. Final anti-RAD17 antibody concentration is 1 mg/ml in PBS buffer. For longer periods of storage, store at -20°C. Avoid repeat freeze-thaw cycles.

Product Protocols: RAD17 antibody tested with Human Ovcar-3 Cells (ARP30546_P050)

Aviva Systems Biology is the original manufacturer of this RAD17 antibody (ARP30546_P050)

Click here to view the RAD17 antibody Western Blot Protocol

Product Datasheet Link: RAD17 antibody (ARP30546_P050)

WB Suggested Anti-RAD17 Antibody Titration: 0.2-1 ug/ml
ELISA Titer: 1:62500
Positive Control: OVCAR-3

Western Blot image:


Description of Target: RAD17 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.

Questions pertaining to this data can be directed to techsupport@avivasysbio.com

Aviva Systems Biology’s RAD17 antibody (ARP30546_P050) has been tested using other cell lysates and tissues. To obtain more data about this antibody please email us at info@avivasysbio.com.

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All of Aviva’s products are guaranteed for the applications and experimental sample types mentioned in the datasheet below. Are you curious if this product will work for you? Please contact us at techsupport@avivasysbio.com

Product Review: RAD17 antibody -N-terminal region (ARP30546_P050) in HeLa lysate, Xenopus laevis egg extract, mouse embryonic stem cell and HEK293T lysate using Western Blot

Product Page for RAD17 antibody-N-terminal region (ARP30546_P050)

Application:Western blotting
Researcher:Domenico Maiorano, Institute of Human Genetics, CNRS
Species+tissue/cell type:
Lane1: 25ug HeLa lysate
Lane2: 25ug Xenopus laevis egg extract
Lane3: 25ug mouse embryonic stem cell lysate
Lane4: 25ug HEK293T lysate
Primary antibody dilution:1:500
Secondary antibody:Anti-rabbit-HRP
Secondary antibody dilution:1:3000

Questionnaire:
Would you use this antibody in future experiment?	YES
Have you used another antibody which has worked in your application?	YES
Do you believe the information about the reagent on Aviva’s website is correct?	YES
If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?	YES because it recognizes the corresponding protein.
How did you store the antibody after re-suspension?	4°C
Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):	1) H. sapiens, M. musculus, X. laevis; 2) Hela cell lysate, mouse embryonic stem cells lysate, Xenopus egg extracts; 3) 25 ug protein per lane.
How many different experimental trials were conducted using the antibody sample?	5
How was this sample prepared?	Hela cell and mouse embryonic stem cells lysates were prpared by lysing  cells direct in into Laemmli buffer followed by boiling. Xenopus egg extracts were prepared by low speed (10 000 g) centrifugation of activated layed eggs, followed by inactivation in Laemmli buffer and boiling.
Primary antibody dilution and incubation time:	1:500 in PBS/0.1% Twenn-20, incubated over night at 4°C.
Secondary antibody used and dilution and incubation time:	1:3000 in 5% non-fat milk in PBS/0.1% Tween -20
What controls were used in your experiment (positive/negative)?	Chromatin fractions isolated from nuclei formed in Xenopus egg extracts irradiated (+) or not (-) with UV light. In these conditions Rad17 is expected to bind to chromatin (positive control ) and not to bind in the absence of aphidicolin (negative control). Histone H3 was used as a chromatin loading control and RPA32 as a positive control of UV irradiation (appearance of slow migrating phosphorylated forms) .
Please include your detailed WB Procedure/Protocol here:	Proteins fractionated on 10% SDS-PAGE were transferred to nitrocellulose membrane (Protran, Whatmann) for 1 hour at 4°C in Tris-glycine buffer supplemented with 20% Ethanol. Membranes were blocked for 1 hour at room temperature with  5% non-fat milk in PBS/0.1% Tween -20. Membranes were incubated over night with primary antibody at 4°C on a shaker. The day after membranes were washed three time for 10 minutes each in PBS/0.1% Tween -20 at room temperature. Membranes were incubated with secondary antibodies in 5% non-fat milk in PBS/0.1% Tween -20 for 4 hours at room temperature on a shaker, then membranes were washed as above and revealed  by ECL (illuminata, crescendo, Millipore).