Description
Hemoglobin Assay Kit (Colorimetric) (BA0043) (BA0043)
The Hemoglobin Assay Kit (Colorimetric) (SKU: BA0043) provides a simple, direct and automation-ready method for measuring total haemoglobin in serum, plasma and urine. Haemoglobin is made of four globin chains, each carrying a haem group, and is carried by red blood cells to transport oxygen from the lungs to peripheral tissues. Quantitation of blood haemoglobin has long been a key diagnostic parameter for conditions such as anaemia, polycythaemia and dehydration. The kit is based on an improved aqueous alkaline method in which haemoglobin is converted into a uniform coloured end product, with the colour intensity measured at 400nm being directly proportional to the haemoglobin concentration. The optimised formulation exhibits high sensitivity and is ideal for measuring haemolysis in low-haemoglobin samples such as serum and plasma. The mix-and-read procedure can be readily automated for high-throughput screening.
| Product Name: | Hemoglobin Assay Kit (Colorimetric) (BA0043) |
| SKU: | BA0043 |
| Detection Method: | Colorimetric |
| Detection Range: | 0.9 - 200 mg/dL haemoglobin in 96-well plate assay |
| Sample Type: | Serum, plasma, urine |
| Species Reactivity: | All |
| Assay Time: | 5 minutes |
| Kit Size: | 250 Assays |
| Equipment Required: | Microplate reader |
| Storage: | The kit is shipped at room temperature. Store reagent and calibrator at 4°C. |
| Shelf Life: | 12 months after receipt |
| Shipping: | Room Temperature |
This kit is based on an improved aqueous alkaline method in which haemoglobin is converted into a uniform coloured end product. The intensity of the colour, measured at 400nm, is directly proportional to haemoglobin concentration in the sample.
- Sensitive and accurate: linear detection range 0.9 - 200 mg/dL haemoglobin in 96-well plate assay
- Simple and high-throughput: the mix-and-read procedure adds a single working reagent and reads the optical density, readily automated for thousands of samples per day
- Safety: reagents are non-toxic
- Versatility: assays can be executed in 96-well plate or cuvette
- Direct assays: total haemoglobin in serum, plasma, urine, etc.
- Pharmacology: effects of drugs on haemoglobin metabolism
- Drug discovery: HTS for drugs that modulate haemoglobin levels
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Blank and Calibrator: pipette 50 µL water (Blank) and 50 µL Calibrator into wells of a clear bottom 96-well plate, then transfer 200 µL water into the Blank and Calibrator wells (the diluted calibrator is equivalent to 100 mg/dL haemoglobin). |
| 2 | Samples: serum and plasma can be assayed directly (n = 1). Transfer 50 µL samples into wells, avoiding bubble formation, add 200 µL Reagent and tap plate lightly to mix. |
| 3 | Incubate 5 minutes at room temperature and read OD at 390-405nm (peak 400nm). |
Subtract blank OD (water) from the Calibrator and Sample OD values. Haemoglobin concentration = [(ODSAMPLE − ODBLANK) / (ODCALIBRATOR − ODBLANK)] × 100 × n (mg/dL), where 100 mg/dL is the equivalent haemoglobin concentration of the diluted calibrator and n is the dilution factor. Conversion: 1 mg/dL Hb equals 0.156 µM, 0.001% or 10 ppm.
| Component | Quantity | Storage |
| HB Reagent | 50 mL | 4°C |
| Calibrator | 10 mL | 4°C |