Product Name: ATR-pS435+pS436
Product Number: AB-PK528
Size: 25 µg      Price:89.00
      $US
Target Full Name: Ataxia telangiectasia and Rad3 related protein-serine kinase

Target Alias: FRAP-related protein; FRP1; MEC1; SCKL; SCKL1; CCDS3124.1; ENSG00000175054 SCKL1;

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK528

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: S435+S436

Protein UniProt: Q13535

Protein SigNET: Q13535

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human ATR sequence peptide Cat. No.: PE-04AHM99
Antibody Immunogen Sequence: RRL(pS)(pS)SLN(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues R432 to N439; In the N-terminal quarter of the kinase after the transmembrane domain. These are This is the major in vivo phosphorylation sites in ATR.

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Antibody Concentration: 1 mg/ml

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 2 µg/ml for immunoblotting

Antibody Potency: Strong immunoreactivity with immunogen peptide on dot blots.

Antibody Species Reactivity: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 270-300 kDa.

Antibody Specificity: High-very high

Antibody Cross Reactivity: Strong 140 kDa protein detected in HEK-293 cells and sea star oocytes. A very strong 38 kDa cross-reactive protein was detected in sea star oocytes.

Related Product 1: ATR-pS435+pS436 blocking peptide

Related Product 2: ATR-2 pan-specific antibody (Cat. No.: AB-NK237-1)

Related Product 3: ATR-3 pan-specific antibody (Cat. No.: AB-NK237-2)

Related Product 4: ATR-4 pan-specific antibody (Cat. No.: AB-NK237-3)

Scientific Background: ATR is a protein-serine/threonine kinase of the Atypical group and PIKK family. It functions as a DNA damage sensor, and plays a pivotal role in the regulation of the cell cycle. ATR appears to be a tumour suppressor protein; cancer-related mutations in human tumours point to a loss of function of the protein kinase. Active ATR normally acts to inhibit tumour cell proliferation. ATR levels are up-regulated 1.6-fold in human tumours compared to most other protein kinases. Loss-of-function mutations in ATR, which abolish its DNA damage detection ability, have been linked to hemangiomas. Somatic mutations in the ATR gene are rarely observed in human cancer specimens, with the possible exception of sporadic stomach and endometrial cancers that display microsatellite instability. In animal studies, mice heterozygous for a loss-of-function mutation in the ATR gene exhibit similar survival times as wild-type mice, but have an increased occurence of tumour formation. Caffeine is an inhibitor of ATR. Interestingly, caffeine exposure is known to sensitize tumours to ionizing radiation and other toxic agents, which is associated with the disruption of cell-cycle checkpoints. It works in a complex with ATRIP. ATR responds to DNA damage and replication stresses and activates proteins that mediate DNA repair. It's other roles involve checkpoint signalling by interacting with CHK1, fragile site stabilty and regulation of centrosome duplication. Upon cellular exposure to genotoxic stresses, such as ionizing radiation (IR), ultraviolent light (UV), or DNA replication stalling, ATR phosphorylates BRCA1, CHEK1, MCM2, RAD17, RPA2, SMC1, and p53/TP53, which collectively function to promote cell cycle arrest, DNA repair, recombination, and potentially apoptosis.
Figure 1. Epitope mapping of ATR-pS435+pS436 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to ATR-pS435+pS436.