IT InnoZyme Calpain 1/2 Activi 1KIT

IT InnoZyme Calpain 1/2 Activi 1KIT

Stock Code: 3586717
Manufacturer Part No: CBA054-1KIT
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Analysis Note


Positive ControlCalpain 1


Components


96-Well Plate, Calpain 1 Control, Substrate, Assay Buffer, Activation Buffer, Inhibitor, Reducing Agent, Plate Sealer and a user protocol.


General description


A highly sensitive and specific assay for measuring Calpain 1 and 2 activity in biological samples like cell lysate or tissue extracts and for screening enzyme inhibitors. Calpains (calcium ion-dependent papain-like cysteine proteases) are widely distributed in all mammalian cells. They mediate calcium-induced activation of the Erk1/Erk2 MAPK pathway and are believed to participate in intracellular signal processing via limited proteolysis of their targets. Sensitive: detects calpain activity at 63-1000 ng/ml Specific: Substrate and inhibitor optimized for specific detection of calpain activity Versatile: Homogeneous 96-well format can be used with biological samples or for inhibitor screens Convenient: Contains all reagents to assay samples in 30 min


Legal Information


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


Other Notes


Higuchi, M., et al. 2005. J. Biol. Chem.280, 15229;Tompa, P., et al. 2004. J. Biol. Chem.279, 20775;Veeranna, et al. 2004. Am. J. Pathol.165, 795;Dainese, E., et al. 2002. J. Biol. Chem.277, 40296;Wang, K.K., et al. 2000. Trends Neurosci.23, 20;Black, R.A., et al. 1997. Nature385, 729;Molinari, M. and Carafoli, E. 1997. J. Membr. Biol.156, 1;Sorimachi, H., et al. 1997. Biochem. J.328, 721.


Packaging


1 kit in Glass bottle


Preparation Note


All reagents necessary to perform the assay are supplied with the kit. Warm all reagents (except Calpain Control and Assay Buffer) to room temperature (15-25°C) just prior to use. The following reagent preparation guidelines will result in sufficient volume for one 96-well plate. Control, Human Calpain 1, diluted: Dilute the Control, Human Calpain 1 100-400-fold with chilled Assay Buffer; maintain on ice at all times. Activation Buffer with Reducing Agent: Add 80 µl Reducing Agent to 9.92 ml Activation Buffer. Substrate (1X): Dilute Substrate 1:25 by adding 200 µl Substrate to 4.8 ml Assay Buffer Inhibitor (1X): Dilute the Inhibitor as follows:1. Dilute the Inhibitor 1:50 by adding 2 µl Inhibitor to 98 µl Assay Buffer2. Add 50 µl of the 1:50 dilution (step 1) to 4.95 ml Activation Buffer with Reducing Agent.


Dilute samples with Assay Buffer as needed.Note: Calpain is a temperature sensitive enzyme. Dilutions of calpain control must be prepared using chilled Assay Buffer and kept on ice at all times.Guidelines for sample preparation: The recommended protein concentration for cell lysates is ≥2 mg/ml as determined using a standard protein assay (e.g. Non-Interfering Protein Assay™, Cat. No. 488250 or BCA Protein Assay, Cat. No. 71285)

Quality Level100
ManufacturerSIGMA-ALDRICH

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