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| |  | Cat. No. DR1036 |
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| Host: Rabbit | | Isotype: IgG | | Immunogen: a recombinant protein consisting of full-length, human Chk2 fused to a His•Tag® sequence and expressed in insect cells | | Form: Liquid | | Formulation: Undiluted serum. | | Preservative: ≤0.1% sodium azide | | Positive Control: HeLa cells | | Comments: Recognizes the ~62 kDa Chk2 protein in HeLa cells. | | Ref.: Chan, D.W., et al. 2000. J. Biol. Chem. 275, 7803. Lee, J.S., et al. 2000. Nature 404, 201. Blasina, A., et al. 1999. Curr. Biol. 9, 1. Tominaga, K., et al. 1999. J. Biol. Chem. 274, 31463. Sanchez, Y., et al. 1997. Science 277, 1497. |
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Lane 1: Detection of human Chk2 by immunoblotting. Sample: Whole cell lysate (50 mg) from HeLa cells. Primary antibody: Anti-Chk2 Rabbit pAb (Cat. No. DR1036) (1:10,000). Detection: chemiluminescence. Lane 2: Detection of human Chk2 by immunoprecipitation followed by immunoblotting. Sample: Whole cell lysate from HeLa cells. Antibody for IP: Anti-Chk2 Rabbit pAb (Cat. No. DR1036) (5 mg/500 mg total protein). Primary antibody: Anti-Chk2 (Ab-2) Mouse mAb (2CHK01) (Cat. No. CC44) (1 mg/ml). Detection: chemiluminescence.
Detection of human Chk2 activity by IP activity assay. Near-confluent HeLa cells were left untreated (-CPT) or treated with camptothecin (+CPT) (10 mM for 2 h). Cell lysates were prepared using 0.5 ml PhosphoSafe™ Extraction Reagent (Cat. No. 71296) per 10 cm culture dish according to the recommended protocol. Equal amounts of total protein (500 mg) were immunoprecipitated using 5 ml Anti-Chk2 Rabbit pAb (Cat. No. DR1036) or 5 ml pre-immune serum. The activity was determined using the K-LISA™ Checkpoint Activity Kit (Cat. No. CBA020) according to the recommended protocol.
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