UN 3316 9
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
Yes, but you would need to have a plate reader that has a kinetic function as this is a kinetic assay.
The cells must be lysed before assaying with this kit. The best way to perform lysis is to incubate the cells with hypotonic buffer (at least 2 volumes and better with 3-4 volumes, depending on the expected level of peroxidase in these cells). Let them swell and homogenize with a Dounce homogenizer. Centrifuge the homogenate at 10,000 × g and use the supernatant for the assay. If a detergent is used for lysis, only use a peroxide-free detergent and follow the instructions in the Bulletin: "Nonionic detergents such as TWEEN and TRITON X-100 that contain high levels of endogenous peroxides will raise the apparent activity. If these detergents are vital to the extraction of the proteins of interest, a low peroxide detergent should be used such as Product Codes X-100-PC (TRITON X-100), P 6585 (TWEEN 20), or P 8192 (TWEEN 80)."
We have not tested the CGP1 assay with tissue samples. However, we suggest that the customer perform a small-scale pilot experiment using our CelLytic-MT lysis buffer (Product No. C3228). The buffer enables efficient extraction of tissue proteins. It does not contain any of the interfering agents that are listed on the kit bulletin.
There is a decrease in absorbance during the assay. The OD values decrease with the increase in reaction time. The slope of the reduction in OD (delta OD per minute) is the value used to determine the enzyme units.
In general, when human erythrocytes were assayed for the enzyme, the sample was diluted 10-fold (15 mg peroxidase per mL) and a 20 μl sample gave a decrease of 0.032 OD340 per minute. Too much enzyme will underestimate the apparent activity of the sample. When rabbit reticulocytes were assayed at a 10-fold dilution (10 mg peroxidase per mL), 20 μl gave a decrease of 0.065 OD340 per minute.
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