Catalase Assay Kit

Stock Code: 3586488
Manufacturer Part No: 219265-1KIT

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Descriptions

Analysis Note

Calculating the Resultsa. Calculate the average absorbances of each standard and sample.b. Subtract the average absorbance of standard A from itself and all other standards and samples.c. Plot the corrected absorbance of standards (from step 2 above) as a function of final formaldehyde concentration (M) from Table 1. See Figure 1 (below) for a typical standard curve.﹤div class="Bio_doc_image">Figure 2: Typical Formaldehyde Standard Curve.﹤/div>d. Calculate the formaldehyde concentration of the samples using the equation obtained from the linear regression of the standard curve substituting corrected absorbance values for each sample.﹤div class="Bio_doc_image">Figure 3: Formaldehyde Concentration Equation﹤/div>e. Calculate the catalase activity of the sample using the following equation. One unit is defined as the amount of enzyme that will cause the formation of 1.0 nmol formaldehyde per min at 25°C.﹤div class="Bio_doc_image">Figure 4: Activity Equation﹤/div>

Components

Assay Buffer, Sample Buffer, Formaldehyde Standard, Catalase, Potassium Hydroxide, Methanol, Hydrogen Peroxide, Purpald (Chromogen), Potassium Periodate, 96 Well Plate, Plate Cover

General description

A sensitive spectrophotometric assay (540 nm) for the measurement of catalase (CAT) activity in plasma, serum, erythrocyte lysates, tissue homogenates, and cell lysates. Assay range: 0.25-4 nmol/min/ml. Contains sufficient reagents for 96 tests.

Legal Information

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

Other Notes

Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.

Wheeler, C.R., et al. 1990. Anal. Biochem.184, 193;Johansson, L.H., et al. 1988. Anal. Biochem.174, 331.

Packaging

1 kit in Glass bottle

Preparation Note

Note: Overheating can inactivate catalase. The enzyme should be kept cold during sample preparation and assaying. In general, catalase is very unstable at high dilution. It is recommended to store samples concentrated and assay within 30 min after dilution.A. Tissue Homogenates1. Prior to dissection, either perfuse tissue or rinse tissue with a phosphate buffered saline (PBS) solution, pH 7.4, to remove any red blood cells and clots.2. Homogenize the tissue on ice in 5-10 ml cold buffer (i.e., 50 mM potassium phosphate, pH 7.0, containing 1 mM EDTA) per gram tissue.3. Centrifuge at 10,000 x g for 15 min at 4°C.4. Remove the supernatant for assay and store on ice. If not assaying on the same day, freeze the sample at -80°C. The sample will be stable for at least one month.B. Cell Lysates1. Collect cells by centrifugation (i.e., 1000-2000 x g for 10 min at 4°C). For adherent cells, do not harvest using proteolytic enzymes; rather use a rubber policeman.2. Homogenize or sonicate the cell pellet on ice in 1-2 ml cold buffer (i.e., 50 mM potassium phosphate, pH 7.0, containing 1 mM EDTA).3