Drabkin’s Microplate Hemoglobin Assay
Drabkin’s Microplate Hemoglobin Assay is for the determination of hemoglobin in blood samples, column fractions, and any sample with visible hemoglobin. It is for research purposes and is not intended for diagnostic use.
Description: The Drabkin’s Microplate Hemoglobin Assay converts the variable hemoglobin forms in blood samples to cyanmethemoglobin which can be read spectrophotometrically at 540 nm. Compared to the standard cuvette-based assay, the microplate assay uses less sample and is amenable to high-throughput. The dose response of the assay is linear from 0.5 mg/ml to 20 mg/ml.
Background: Hemoglobin can exist in multiple forms depending on oxygen content, carbon monoxide content, and oxidative state of the iron (ferrous, ferric, and ferryl). Each form has a unique extinction coefficient and peak absorbance wavelength. The heterogeneous nature of hemoglobin can be overcome by conversion to cyanmethemoglobin. This is achieved by reaction of hemoglobin with alkaline ferricyanide and cyanide in a single reagent. The resulting cyanmethemoglobin has a peak absorbance at 540 nm and an extinction coefficient of 0.68 for a 1 mg/ml solution.
- 1 x 96-well Assay Plate
- 1 x Hemoglobin Standard
- 2 x Drabkin’s Reagent
Other Materials required but not provided:
- Microplate reader equipped to measure absorbance at 540 nm is required
- Adjustable pipettes and pipette tips
- Multi-channel pipettor(s) and tips for dispensing 150 µl volumes
- Tubes for preparing dilutions, i.e. microfuge tubes
|Dimensions||11 × 10 × 15 in|