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Lactate dehydrogenase (LD or LDH) catalyzes the reduction of pyruvate by NADH to form lactate and NAD+. The catalytic concentration is determined from the rate of decrease of NADH measured at 340 nm.
PREPARATION OF WORKING SOLUTION
Prepare working solution by mixing Reagent R1 and Reagent R2 in the ratio 4:1 as per requirement.
REAGENT STORAGE & STABILITY
Tris buffer, pH 6.8
SPECIMEN COLLECTION & PRESERVATION
Collect the specimen in a clean & dry container. Although serum is preferred, plasma with Heparin or EDTA can be used. Hemolyzed samples should not be used since LDH activity in erythrocytes is 160 fold higher than in serum. The serum should be separated from the clot promptly. Samples should be assayed soon after collection. LDH is stable in serum or plasma for four days at 2 - 8 degree C. Do not freeze or expose the serum to high temperature as this may inactivate thermolabile LDH isoenzymes.
Prewarm at 37 degree C the required amount of working solution before use. Perform the assay as given below :
1.0 ml procedure
Mix and aspirate. After the initial delay of 60 seconds, record the absorbance of the test at an interval of 30 seconds for the next 90 seconds at 340nm.
25 degree C
30 degree C
37 degree C
240 - 480
4.00 - 8.00
The following factors are used for conversion: