Invitrogen™ Hemoglobin Colorimetric Detection Kit

The Hemoglobin Detection research-use-only kit is a colorimetric assay designed for the quantification and detection of hemoglobin in Whole Blood, RBCs, and hemolyzed serum and plasma samples.

This complete, ready-to-use kit includes clear 96-well plate(s), hemoglobin standard, and other components to perform the assay. A 96-well microplate reader capable of reading optical density between 560 and 580 nm is required for use of this kit.

Performance characteristics
• Assay type: colorimetric detection kit
• Sample types: whole blood, RBCs, and hemolyzed serum and plasma samples
• Sensitivity 0.021 g/dL
• Standard curve range: 0.25–16 g/dL
• Reactivity: species independent

Background
Hemoglobin (Hgb) is an erythrocyte protein complex comprised of two sets of identical pairs of subunits, each of which bind an iron-prophyrin group commonly called heme. Generally containing two alpha or alpha-like globulin chains, the remaining subunits may be beta, gamma, delta or epsilon, or in the case of infants, fetal hemoglobin that is replaced during the first year of life. Heme binds and releases oxygen or carbon dioxide in response to slight changes in local gas tension. Free oxygen or carbon dioxide bound by one heme group facilitates subsequent binding by the other heme groups in a given hemoglobin molecule.

Assay principle
The Hemoglobin Colorimetric Detection Kit is designed to quantitatively measure all forms of hemoglobin present in blood and RBCs, or plasma and serum. A human hemoglobin standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate and the ready-to-use Hemoglobin Detection Reagent is added to each well. For whole blood or RBC samples, 10 μL of samples and standards are used in the Regular format. Results are calculated as g/dL for whole blood and RBCs, and mg/mL for serum and plasma. The plate is incubated for 30 minutes at room temperature. The plate is read at 560-580 nm to detect the intensity of the color generated. This Hemoglobin Detection kit uses a single reaction solution that is light stable at 4°C and does not contain dangerous chemicals. All forms of hemoglobin are rapidly converted to a single stable form that is measured photometrically. Many samples can be measured without dilution in this safe, simple assay.