Hemoglobin antibodies for flow cytometry
We offer antibodies to detect various types of hemoglobin. Each antibody can be used individually or combined in a panel with anti-HbS FITC, anti-panHb APC and anti-HbF Dy-410 for three lasers, eliminating the need for compensation.
Anti-hemoglobin S FITC
This is the first anti-hemoglobin S (HbS) antibody conjugate available for flow cytometry. For convenience, it’s conjugated with FITC, so it can be measured on all available flow cytometers.
The anti-HbS antibody recognizes the abnormal β-chain of HbS. Substitution of the amino acid valin for glutamic acid in the sixth position of the β-globin chain results in the formation of a βs subunit. Due to the mutated subunit βs, hemoglobin becomes unstable when no oxygen is bound, resulting in sickle shaped erythrocytes.
Anti-pan hemoglobin APC
The murine pan hemoglobin (panHb) antibody recognizes the α-chain of human hemoglobin. This antibody can be combined with other antibodies to determine the percentage of erythrocytes containing pathological forms of hemoglobin compared to the total erythrocyte population.
Figure 2. Flow cytometry analysis of whole blood stained with anti-pan hemoglobin. Histograms: uninterrupted line represents stained sample, interrupted line represents blanc sample.
Anti-hemoglobin F Dy-410
The anti-hemoglobin F antibody recognizes the γ-chain of hemoglobin F (HbF). HbF is predominantly expressed by fetal erythrocytes and in some cases, HbF is also expressed by adult erythrocytes. These cells, so called F-cells, can represent up to 25 % of the total erythrocyte population.
Figure 3. Flow cytometry analysis of whole blood stained with anti-hemoglobin F. Histograms: uninterrupted line represents stained sample, interrupted line represents blanc sample.
Complementary permeabilization reagent
IQ Perm
IQ Perm can be used as a permeabilization buffer when working with the above mentioned antibodies. Detection of these intracellular antigens or substrates often require rapid fixation, usually by formaldehyde or glutaraldehyde, followed by treatment with a permeabilizing reagent.
IQ Perm is such a permeabilizing reagent which utilizes Triton X‐100 as the major active component that allows entry of antibodies and other probes. IQ Perm is suitable for permeabilization of erythrocytes and leukocytes. Our experience has shown that IQ Perm is used at various dilutions, depending on the cells of interest.
IQ Perm is provided as a concentrate in 40 mL volume. This bottle is sufficient for 105 ‐ 265 individual tests, depending upon the application using approximately one million cells for each assay.
Details on the available antibodies are listed here and details on IQ Perm can be found here