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Anti-Yeast Rad9 pS1260 (RABBIT) Antibody
Rad9 phospho S1260 Antibody
- SPECIFICATION
- CITATIONS
- PROTOCOLS
- BACKGROUND
| Host | Rabbit |
|---|---|
| Conjugate | Unconjugated |
| Target Species | Yeast |
| Reactivity | Yeast |
| Clonality | Polyclonal |
Application 
| WB, E, I, LCI |
| Application Note | This phospho specific polyclonal antibody was tested by immunoblotting and ELISA. Data from both immunoblotting and ELISA indicate the antibody is reactive with the phosphorylated form of the immunizing peptide and minimally reactive with the non-phosphorylated form of the immunizing peptide. Immunoblotting detects yeast Rad9 protein. No reactivity is expected against the human or mouse analogs of RAD9. Reactivity against RAD9 from other sources is unknown. Cross reactivity may occur with auto-phosphorylated Rad53 kinase. Although not tested, this antibody is likely functional by IHC and IP. This product has been assayed against 0.1 µg of phosphorylated peptide (pS1260) in a standard capture ELISA using TMB (3,3',5,5'-Tetramethylbenizidine) code # TMBE-100 as a substrate for 30 minutes at room temperature. A working dilution of 1:5,000 is suggested for this product. Minimal reactivity was detected against the non-phosphorylated form (S1260) of the immunizing peptide. This antibody appears to be specific for the active form (phosphorylated) of the protein. Dilute the antibody 1:100 to 1:500 for immunoblotting. Researchers should determine optimal titers for other applications. |
| Physical State | Liquid (sterile filtered) |
| Buffer | 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Immunogen | This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to phosphorylated form of an internal region near aa 1225-1275 from the aa1309 yeast Rad9 protein conjugated to KLH. |
| Preservative | 0.01% (w/v) Sodium Azide |
| Gene ID | 851803 |
|---|---|
| Other Names | 851803 |
| Purity | This affinity purified antibody is directed against the phosphorylated form of yeast Rad9 at the pS1260 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody is phospho specific to yeast pS1260. |
| Storage Condition | Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. |
| Precautions Note | This product is for research use only and is not intended for therapeutic or diagnostic applications. |
| Name | RAD9 |
|---|---|
| Function | Essential for cell cycle arrest at the G2 stage following DNA damage by X-irradiation or inactivation of DNA ligase. |
| Cellular Location | Nucleus. |
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Provided below are standard protocols that you may find useful for product applications.
Background
Rad9 is required for the MEC1/TEL1-dependent activation of Saccharomyces cerevisiae DNA damage checkpoint pathways mediated by Rad53 and Chk1. DNA damage induces Rad9 phosphorylation, and Rad53 specifically associates with phosphorylated Rad9. Cells have evolved multiple strategies for tolerating genomic damage. The most important of these are numerous repair systems that remove or bypass potentially mutagenic DNA lesions. Another cellular strategy is to delay cell-cycle transitions at multiple points. The genetic control of these delays, termed `checkpoints', was first established in budding yeast where it was shown that the RAD9 gene functions in G2/M arrest after irradiation with X-rays. Subsequently, it has become clear that Rad9 also functions at the G1/S, intra-S and mid-anaphase checkpoints. Defects in checkpoint regulation can lead to genome instability and, in higher eukaryotes, neoplastic transformation. Rad9 also controls the transcriptional induction of a DNA damage regulon (DDR). Rad9 may also have a pro-apoptotic function. This is suggested in that Rad9 from Schizosaccharomyces pombe (SpRad9) contains a group of amino acids with similarity to the Bcl-2 homology 3 death domain, which is required for SpRad9 interaction with human Bcl-2 and apoptosis induction in human cells. Overexpression of Bcl-2 in S. pombe inhibits cell growth independently of rad9, but enhances resistance of rad9-null cells to methyl methanesulfonate, ultraviolet and ionizing radiation. Rad9 conveys the checkpoint signal by activating Rad53p and Chk1p; is hyperphos-phorylated by Mec1p and Tel1p; and is a potential Cdc28p substrate. Mature yeast Rad9 is reported to have an apparent molecular weight of ~148kDa. The human homolog is reported at 48.5 kDa.
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