Pyruvate Dehydrogenase Beta (PDHb) BioAssay™ ELISA Kit (Human)

Sample Type:
Tissue homogenates, cell lysates, cell culture supernates and other biological fluids

Intended Use:
The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of PDHb in human tissue homogenates, cell lysates, cell culture supernates and other biological fluids.

Sensitivity:
0.65ng/ml

Range:
1.56-100ng/ml

Specificity:
This assay has high sensitivity and excellent specificity for detection of Pyruvate Dehydrogenase Beta (PDHb). No significant cross-reactivity or interference between Pyruvate Dehydrogenase Beta (PDHb) and analogues was observed.

Precision:
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Test Principle:
The microplate provided in this kit has been pre-coated with an antibody specific to PDHb. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to PDHb. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain PDHb, biotin-conjugated antibody and
enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of PDHb in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Kit Components:
96-well strip plate, 1x Plate
Standard, 2x vial
Detection Reagent A, 1×120ul
Detection Reagent B, 1×120ul
TMB Substrate, 1×9ml
Wash Buffer (30X), 1×20ml
Standard Diluent, 1×20ml
Assay Diluent A, 1×12ml
Assay Diluent B, 1×12ml
Stop Solution, 1x6ml

Storage and Stability:
Store microtiter strips, Standard, Detection Reagent A, Detection Reagent B and positive control at -20°C. After reconstitution of the postive control store at 4°C and use within 5 days. Store all the other components at 4°C. Unused kit is stable for 6 months after receipt. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.