Here is the recommended protocol for cell sample preparation:
1. Homogenize cells with the heme reagent which contains NaOH and detergent which will effectively lyse the cells.
2. Let incubate for 5 min for the color to develop, then centrifuge to pellet any debris.
3. Transfer 250 µL of clear supernatant to a 96 well plate and measure the absorbance at 400 nm.
MAK316
Heme Assay Kit
sufficient for 250 colorimetric tests
Synonym(s):
Heme Quantification Kit
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About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.
Recommended Products
detection method
colorimetric
relevant disease(s)
hematological disorder
storage temp.
2-8°C
General description
Heme is a member of the porphyrin family. It is synthesized in both mitochondria and cytoplasm, and is a key prosthetic group for various essential proteins such as hemoglobin, cytochromes, catalases and peroxidases. Free heme, which can be released from hemoglobin following hemolysis, is pro-inflammatory and contributes to iron-derived reactive oxygen species. Heme determination is widely practiced by researchers of various blood diseases.[1][2]
Features and Benefits
Compatible with high-throughput handling systems.
Suitability
Suitable for the quantitative determination of heme and evaluation of drug effects on heme metabolism in a variety of biological samples such as blood, plasma, serum, urine, and heme carrying enzymes.
Principle
Heme assay is based on an aqueous alkaline solution method, in which the heme is converted into a uniform colored form, producing a colorimetric (400 nm) result, directly proportional to the heme concentration in the sample. The optimized formulation reduces interference and exhibits high sensitivity. One mg/dL heme equals 15.3M, 0.001%, or 10ppm. The kit has a linear detection range of 0.6–125M heme in a 96 well format.
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Aquatic Acute 1 - Aquatic Chronic 2 - Eye Irrit. 2 - Met. Corr. 1
Storage Class Code
8B - Non-combustible corrosive hazardous materials
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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How do you use this product on cell culture sample? How to homogenize the sample prior of heme measurement?
1 answer-
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How can I determine the shelf life / expiration / retest date of this product?
1 answer-
If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/449/386/product-dating-information-mk.pdfHelpful?
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How is shipping temperature determined? And how is it related to the product storage temperature?
1 answer-
Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf
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How does this kit work with cell culture sample? How do you homogenize the cells prior of measurement?
1 answer-
This kit was validated for use with serum, plasma and whole blood. Cell lysates have not been analyzed with this kit and no specific guidance is provided for this type of sample preparation.
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Could I use distilled water instead of ultrapure? Can I use this kit to measure how much plasma free hemoglobin I have in a plasma sample?
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Only ultra pure water should be used because it is filtered through a 0.22 μm filter which removes any bacteria that could interfere with the chemical reaction.
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Does MAK316 work for mouse sera?
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MAK316 is expected to work for all species and should be compatible with mouse sera samples, as well as with other sample types such as blood, serum, plasma, and urine.
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how is this kit used
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The kit is used according to the product protocol. Here is a link to that document:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/386/102/mak316bul.pdf
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お世話になります。臨床研究でヒトの血栓を回収してホルマリン固定しております。遊離ヘムを染めたいと考えておりますが 可能でしょうか？また、検体を送れば染めていただけるのでしょうか？
1 answer-
This product is specifically tested for the quantitative determination of heme in various biological samples. Solid tissues are homogenized and lysed by the heme reagent which contains NaOH and a detergent. This would also apply to the clot sample resulting in the lysis of any intact RBCs. Given the principle of the kit, measuring only the free heme would not be possible. Currently, there is no product available to meet your requirements. Unfortunately, lab services are not offered for this particular application.
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