Methaemoglobin assay kit (Colorimetry Method)
Size: 100T/96S
Application: For detection of Methemoglobin content in various animal whole blood and other samples.
Storage and Expiration: Store at 2-8℃ in dark for 6 months.
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Price: ---
Catalog No.: BYEK3811
Size: 100T/96S
Application: For detection of Methemoglobin content in various animal whole blood and other samples.
Storage and Expiration: Store at 2-8℃ in dark for 6 months.
Introduction
Methemoglobin (British: methaemoglobin) (pronounced "met-hemoglobin") is a hemoglobin in the form of metalloprotein, in which the iron in the heme group is in the Fe3+ (ferric) state, not the Fe2+ (ferrous) of normal hemoglobin. Sometimes, it is also referred to as ferrihemoglobin. Methemoglobin cannot bind oxygen, which means it cannot carry oxygen to tissues. It is bluish chocolate-brown in color. In human blood a trace amount of methemoglobin is normally produced spontaneously, but when present in excess the blood becomes abnormally dark bluish brown. The NADH-dependent enzyme methemoglobin reductase (a type of diaphorase) is responsible for converting methemoglobin back to hemoglobin.
Material Required But Not Provided
1.Test tubes
2. Micropipettes
3. Vortex mixers
4. Visible spectrophotometers (540, 602 and 630nm)
Kit Components
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Components |
Storage Instruction |
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1 |
Reagent A |
Hemoglobin Assay Concentrate Solution |
1ml×4 vial, can be stored at 2-8℃. |
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Reagent A working solution preparation: Dilute Reagent A with double-distilled water at ratio of 1:99, can be stored at 2-8℃ in dark for 1 month. |
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2 |
Reagent B |
Methemoglobin Assay Concentrate Solution |
60ml×1 bottle, can be stored at 2~8℃. |
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Reagent B working solution preparation: Dilute Reagent B with double-distilled water at ratio of 1:9, can be stored for 6 month. |
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3 |
Reagent C Powder |
White Powder |
1 vial, can be stored at 2-8℃ in dark |
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Reagent C diluent buffer |
diluent buffer |
1 ml, can be stored at 2-8℃. |
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Reagent C working solution preparation (For making reduction hemoglobin curve): Add 1ml of Reagent C diluent buffer to Reagent C Powder before use, be careful to avoid light. |
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4 |
Reagent D Powder |
Brick red Powder |
1 vial, can be stored at 2-8℃ in dark |
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Reagent D diluent buffer |
diluent buffer |
1 ml, can be stored at 2-8℃. |
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Reagent D working solution preparation (For making methemoglobin curve): Add 1ml of Reagent D diluent buffer to Reagent D Powder before use, be careful to avoid light. |
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Protocol
1. Preparation of Anticoagulated Whole Blood: The whole blood was immediately added to the heparin anticoagulant tube, sealed with a cap, and mixed by gentle inversion.
2. Determination of hemoglobin content: Take 0.01ml of whole blood and 2.5ml of reagent A working solution, mix well, let stand for 5 minutes, 1cm optical path, double-distilled water adjust to zero, and measure the absorbance value of each tube at 540nm.
3. Methemoglobin determination: Take 0.05ml of whole blood, add 2.5ml of reagent B working solution, mix well, let stand for 5 minutes, 1cm optical path, double-distilled water adjust to zero, and measure the absorbance at 630nm and 602nm value. If there is no dual-wavelength spectrophotometer, the absorbance value at 630 nm of each tube can be measured first, and then the absorbance value at 602 nm of each tube can be measured. But be sure to pay attention not to mistake the number of each tube.
