Metabolism Assays
Heme Assay Kit (BA0044)
- SKU:
- BA0044
- Product Type:
- Assay
- Instrument:
- Microplate Reader
- Sample Type:
- Blood, Serum, Plasma, Urine
- Research Area:
- Clinical Chemistry
Description
Heme Assay Kit (BA0044) (BA0044)
The Heme Assay Kit (SKU: BA0044) offers a simple, direct and automation-ready method for measuring total haem in blood, serum, plasma, urine and haem-carrying enzymes. Haem is an important member of the porphyrin family, synthesised in both mitochondria and cytoplasm and serving as a key prosthetic group for essential proteins such as haemoglobin, cytochromes, catalases and peroxidases. Haem determination is widely practised by researchers studying various blood diseases. The kit is based on an improved aqueous alkaline solution method in which haem is converted into a uniform coloured form, with the colour intensity measured at 400nm being directly proportional to the haem concentration. The optimised formulation substantially reduces interference by substances in raw samples and exhibits high sensitivity. The mix-and-read procedure can be readily automated for high-throughput screening.
| Product Name: | Heme Assay Kit (BA0044) |
| SKU: | BA0044 |
| Detection Method: | Colorimetric |
| Detection Range: | 0.6 - 125 µM haem in 96-well plate assay |
| Sample Type: | Blood, serum, plasma, urine, haem-carrying enzymes |
| 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 solution method in which haem is converted into a uniform coloured form. The intensity of the colour, measured at 400nm, is directly proportional to the haem concentration in the sample.
- Sensitive and accurate: linear detection range 0.6 - 125 µM haem 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 haem in blood, serum, plasma, urine and haem-carrying enzymes
- Pharmacology: effects of drugs on haem metabolism
- Drug discovery: HTS for drugs that modulate haem 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 62.5 µM haem). |
| 2 | Samples: serum and plasma can be assayed directly (n = 1); dilute blood samples 100-fold in distilled water (n = 100). 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 380-420nm (peak 400nm). |
Subtract blank OD (water) from the Calibrator and Sample OD values. Total haem concentration = [(ODSAMPLE − ODBLANK) / (ODCALIBRATOR − ODBLANK)] × 62.5 × n (µM), where n is the dilution factor (100 for blood samples). Conversion: 1 mg/dL haem equals 15.3 µM, 0.001% or 10 ppm.
| Component | Quantity | Storage |
| Reagent | 50 mL | 4°C |
| Calibrator (equivalent to 62.5 µM haem) | 10 mL | 4°C |
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