Rad9 Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-77978

Novus Biologicals, part of Bio-Techne
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NBP1-77978
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Key Product Details

Species Reactivity

Validated:

Yeast

Applications

ELISA, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all Rad9 Antibodies »

Product Summary for Rad9 Antibody

Immunogen

This affinity purified Rad9 Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near aa 1120-1145 from the aa1309 yeast Rad9 protein conjugated to KLH. (Uniprot: P14737)

Reactivity Notes

No reactivity is expected against human and mouse homologs. Reactivity to Rad9 from others sources is unknown.

Specificity

This affinity purified antibody is directed against an internal sequence of yeast Rad9 at the S1129 residue. This pan reactive polyclonal antibody reacts equally with both phosphorylated and non- phosphorylated yeast Rad9 at S1129. No reactivity is expected against human and mouse homologs. Reactivity to Rad9 from others sources is unknown.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This affinity purified antibody is directed against an internal sequence of yeast Rad9 at the S1129 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was purified against the immunizing peptide

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for Rad9 Antibody

Western Blot: Rad9 Antibody [NBP1-77978]

Western Blot: Rad9 Antibody [NBP1-77978]

Western Blot: Rad9 Antibody [NBP1-77978] - Used at a 1:200 dilution incubated 8 hr at room temperature to detect Rad9 by Western blot. Lanes were loaded with 50 ng each of recombinant GST fusion protein containing S. cerevisiae Rad9 (aa 991-1309 ~60 kDa) on a 4-20% Criterion gel for SDS-PAGE as follows: 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.

Applications for Rad9 Antibody

Application
Recommended Usage

ELISA

1:5000

Western Blot

1:100-1:500
Application Notes
This pan reactive polyclonal antibody was tested by immunoblotting and ELISA. Data from both immunoblotting and ELISA indicate the antibody is pan reactive with both the phosphorylated and non-phosphorylated forms of the peptide and protein. Immunoblotting detects yeast Rad9 protein. No reactivity is expected against human and mouse homologs. Reactivity to Rad9 from others sources is unknown. Although not tested, this antibody is likely functional by immunohistochemistry and immunoprecipitation. This product has been assayed against 0.1 ug of immunizing peptide (S1129) in a standard capture ELISA using TMB (3,3',5,5'-Tetramethylbenizidine) as a substrate for 30 minutes at room temperature. A working dilution of 1:5,000 is suggested for this product. Reactivity was detected against both the phosphorylated and non-phosphorylated form (S1129 and pS1129) of the immunizing peptide. This antibody appears to be pan reactive for both forms of the protein. Dilute the antibody 1:100 to 1:500 for immunoblotting. Researchers should determine optimal titers for other applications.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

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

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: Rad9

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 hyperphosphorylated 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.

Alternate Names

cell cycle checkpoint control protein RAD9A, DNA repair exonuclease rad9 homolog A, EC 3.1.11.2, hRAD9, RAD9, RAD9 (S. pombe) homolog, RAD9 homolog A (S. pombe)

Gene Symbol

RAD9A

UniProt

Q99638 (Human), P14737 (Yeast)

Additional Rad9 Products

Product Documents for Rad9 Antibody

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Rad9 Antibody

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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