RAD9 (phospho S1260) polyclonal antibody
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More Files
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Western Blot (Recombinant protein)
Immunoblotting of RAD9 (phospho S1260) polyclonal antibody (Cat # PAB11310) to yeast RAD9 at pS1260 was usedat a 1 : 200 dilution incubated overnight at 4°C to detect RAD9 by Western blot.
Lanes were loaded with 50 ng each of recombinant GST fusion protein containing S. cerevisiae RAD9 (amino acids 991-1309, ~60 kDa) on a 4-20% Criterion gel for SDS-PAGE.
Lane 1, non-phosphorylated wild type yeast RAD9.
Lane 2, in vitro phosphorylated wild type yeast RAD9.
Lane 3, non-phosphorylated S1129A/S1260A double mutant RAD9.
Lane 4, in vitro phosphorylated S1129A/S1260A double mutant. Phosphorylation of RAD9 was by treatment with ATP and RAD53 kinase.
RAD53 kinase autophosphorylates and appears cross reactive as it is detected on the blot as 90 and 110kDa bands (asterisk).
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic phosphopeptide of RAD9.
Immunogen
Synthetic phosphopeptide (conjugated with KLH) corresponding to residues surrounding S1260 of Saccharomyces cerevisiae RAD9.
Host
Rabbit
Reactivity
Yeast
Specificity
This phospho specific polyclonal antibody reacts with phosphorylated pS1260 of yeast Rad9. Reactivity with non-phosphorylated yeast Rad9 is minimal by ELISA and immunoblotting. No reactivity is expected against the human or mouse analogs of RAD9. Cross reactivity may occur with auto-phosphorylated Rad53 kinase.
Form
Liquid
Quality Control Testing
Antibody Reactive Against Synthetic Peptide.
Recommend Usage
Sandwich ELISA (1:5000)
Western Blot (1:100-1:500)
The optimal working dilution should be determined by the end user.Storage Buffer
In 20 mM KH2PO4, 150 mM NaCl, pH 7.2 (0.01% sodium azide)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Western Blot (Recombinant protein)
Immunoblotting of RAD9 (phospho S1260) polyclonal antibody (Cat # PAB11310) to yeast RAD9 at pS1260 was usedat a 1 : 200 dilution incubated overnight at 4°C to detect RAD9 by Western blot.
Lanes were loaded with 50 ng each of recombinant GST fusion protein containing S. cerevisiae RAD9 (amino acids 991-1309, ~60 kDa) on a 4-20% Criterion gel for SDS-PAGE.
Lane 1, non-phosphorylated wild type yeast RAD9.
Lane 2, in vitro phosphorylated wild type yeast RAD9.
Lane 3, non-phosphorylated S1129A/S1260A double mutant RAD9.
Lane 4, in vitro phosphorylated S1129A/S1260A double mutant. Phosphorylation of RAD9 was by treatment with ATP and RAD53 kinase.
RAD53 kinase autophosphorylates and appears cross reactive as it is detected on the blot as 90 and 110kDa bands (asterisk).Enzyme-linked Immunoabsorbent Assay
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Gene Info — RAD9
Entrez GeneID
851803Protein Accession#
Locus:11q13.1-q13.2;OMIM603761;GDB5592334;SwissProtQ99638Gene Name
RAD9
Gene Alias
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Gene Description
DNA damage-dependent checkpoint protein, required for cell-cycle arrest in G1/S, intra-S, and G2/M; transmits checkpoint signal by activating Rad53p and Chk1p; hyperphosphorylated by Mec1p and Tel1p; potential Cdc28p substrate
Gene Ontology
HyperlinkGene Summary
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Other Designations
Rad9p
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Publication Reference
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Rad9 phosphorylation sites couple Rad53 to the Saccharomyces cerevisiae DNA damage checkpoint.
Schwartz MF, Duong JK, Sun Z, Morrow JS, Pradhan D, Stern DF.
Molecular Cell 2002 May; 9(5):1055.
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Schizosaccharomyces pombe Rad9 contains a BH3-like region and interacts with the anti-apoptotic protein Bcl-2.
Komatsu K, Hopkins KM, Lieberman HB, Wang H.
FEBS Letters 2000 Sep; 481(2):122.
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Rad53 FHA domain associated with phosphorylated Rad9 in the DNA damage checkpoint.
Sun Z, Hsiao J, Fay DS, Stern DF.
Science 1998 Jul; 281(5374):272.
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Rad9 phosphorylation sites couple Rad53 to the Saccharomyces cerevisiae DNA damage checkpoint.
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