ATP Colorimetric/Fluorometric Assay Kit

ATP Colorimetric/Fluorometric Assay Kit-100 ASSAY PRICE@ 39,000/-

 ATP Assay Protocol:

1. Sample Preparation: Lyse 1 x 106 cells or homogenize tissues (10 mg) in 100 l ATP Assay Buffer. Deproteinize cell lysate or tissue homogenate using Deproteinization Sample Preparation Kit  or 10 kDa Spin Column. Add 2-50 l of sample to a 96-well plate. Adjust the volume to 50 l/well with ATP Assay Buffer. Notes: a. As ATP is labile, for more accurate assays, we recommend using fresh samples. For samples to be assayed at later date, snap freeze samples using liquid N2 or dry ice. b. Tissues samples may contain enzymes that consume ATP rapidly. We suggest quick processing of your samples and deproteinization using Deproteinization Sample Preparation Kit (Cat. # K808). c. For unknown samples, we suggest performing a pilot experiment & testing different sample dilutions to ensure the readings are within the Standard Curve range. d. For samples having background, prepare parallel well(s) containing same amount of sample as in the test well. e. Endogenous compounds may interfere with the reaction. To ensure accurate determination of ATP in the test samples, we recommend spiking samples with a known amount of Standard (300 pmol).

2. Standard Curve Preparation: For the colorimetric assay, dilute 10 l of the ATP Standard with 90 l of dH2O to generate 1 mM ATP standard, mix well. Add 0, 2, 4, 6, 8, 10 l into a series of wells and adjust volume to 50 l/well with ATP Assay Buffer to generate 0, 2, 4, 6, 8, 10 nmol/well of ATP Standard. For the fluorometric assay, further dilute the ATP Standard to 0.01- 0.1 mM with the dH2O (Detection sensitivity is 10-100 fold higher with the fluorometric than with the colorimetric assay). Follow the procedure as for the colorimetric assay.  

3. Reaction Mix:Mix enough reagent for the number of samples and standards to be performed: For each well, prepare a total 50 l Reaction Mix