Analysis Note

ControlFor 05-649SP use Jurkat cell lysate, HeLa cell lysate.For 05-740SP use Irradiated HeLa cell lysates.For 05-513SP use Raji cells

Application

Research CategorySignaling

Research Sub CategoryCell Cycle, DNA Replication & Repair

This Antibody pack contains Anti-ATM Antibody, Anti-phospho-ATM Antibody (Ser1981), Anti-Chk2 Antibody validated for use in WB, ICC & IP.

Components

05-513SP Anti-ATM, clone AM905-740SP Anti-phospho-ATM (Ser1981),
clone 10H11.E1205-649SP Anti-Chk2, clone 7

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Pathway Explorer Antibody MiniPack:
Each Pathway Explorer Antibody Minipack contains three related antibodies as part of a signaling cascade or a combination of total and phosphorylated forms of key signaling targets. Each of the three antibodies are 30% the original pack size. Full size versions of each of the Pathway Explorer antibodies are available for sale individually under the same catalog number with the removal of “SP” off of each one (e.g. 05-591SP can be ordered as 05-591).

ATM:
Ataxia telangiectasia mutated kinase (ATM) and ataxia telangiectasia and Rad3-related kinase (ATR) are related kinases that regulate cell cycle checkpoints and DNA repair. Mutation in the ATM gene results in the autosomal recessive disease ataxia telangiectasia (AT). The identified substrates for ATM are p53, p95/NBS1, MDM2, Chk2, BRCA1, CtIP, 4E-BP1 and Chk1. The essential requirement for the substrates of ATM/ATR is S/TQ. Hydrophobic amino acids at positions -3 and -1, and negatively charged amino acids at position +1 are positive determinants for substrate recognition by these kinases. Positively charged residues surrounding the S/TQ are negative determinants for substrate phosphorylation. The complex phenotype of cells derived from patients with AT suggests that ATM has additional cellular substrates. In unirradiated cells, ATM is present as an inactive homodimer or multimer. Double-stranded breaks in DNA caused by ionizing radiation cause rapid ATM kinase activation through dissociation of this complex and ATM autophosphorylation at Ser1981.

Chk2:
The p53 tumor-suppressor gene integrates numerous signals that control cell life and death. Several novel molecules involved in p53 signaling, including Chk2, p53R2, p53AIP1, Noxa, PIDD, and PID/MTA2, were recently discovered. The checkpoint kinase Chk2 is the mammalian homologue of yeast Cds1/Rad53. In response to DNA damage, the checkpoint kinase ATM phosphorylates and activates Chk2, which in turn directly phosphorylates and activates p53. Chk2 serves as ATM downstream effector to mediate activation of p53. Chk2 also phosphorylates and activates BRCA1, the product of a tumor suppressor gene that is mutated in breast and ovarian cancer.

* See full size versions for corresponding references.

Immunogen

Epitope: 05-740SP:Ser1981

05-649SP: Mixture of two GST fusion proteins corresponding to residues 1-223 and 217-543 of human Chk2;05-740SP: KLH-conjugated, synthetic peptide corresponding to amino acids 1974-1988 (SLAFEEG[pS]QSTTISS) of human ATM. The immunizing sequence has 11/12 identical amino acids in mouse and rat.;05-513SP: Full length human ATM. Clone AM9.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Packaging

Each vial is 30% the size of the parent catalog number

Physical form

3 individual tubes containing either Anti-ATM, clone AM9; Anti-phospho-ATM (Ser1981), clone 10H11.E12; or Anti-Chk2, clone 7

Quality

05-649SP: Routinely evaluated by immunoblot on RIPA lysates from Jurkat cells, HeLa, 3T3/A31, or A431 cells.

Western Blot Analysis: 0.1-1 µg/mL of this lot detected Chk2 in RIPA lysates from Jurkat cells. A previous lot detected Chk2 in lysates from HeLa, 3T3/A31, and A431 cells.

05-740SP: Routinely evaluated by immunoblot on in crude lysates from irradiated HeLa cells.

Western Blot Analysis: 0.5 µg/mL of this lot detected phosphorylated ATM in crude lysates from irradiated HeLa cells.

05-513SP: Routinely evaluated by immunoblot on nuclear extract from Raji cells.

Western Blot Analysis:
1:250-1:1000 dilution of this lot detected ATM in a nuclear extract from Raji cells.

Specificity

05-649SP and 05-740SP: Predicted to cross-react with rat based on sequence homology.

05-513SP: Other species not tested.

05-649SP and 05-740SP: Others not tested.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap.

Target description

05-649SP: 67 kDa;05-740SP:~370 kDa;05-513SP: 350 kDa