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Hemin Colorimetric Assay Kit

based on 3 citations in multiple journalsHemin Colorimetric Assay Kit34.1 4
Highly Sensitive Assay
Catalog #: K672
$405.00

Product Details

Cat # +Size K672-100
Size 100 assays
Detection Method Absorbance (570 nm)
Species Reactivity Mammalian
Applications Trace amounts of Hemin can be quantitated in the 5-160 pg (10-250 fmol) range.
Features & Benefits • Simple procedure; takes ~ 30 minutes to complete the assay
• Fast and convenient
• Highly sensitive assays for measuring Heme level in biological samples
Kit Components • Hemin Assay Buffer
• Hemin Probe
• DMSO (anhydrous)
• Enzyme Mix
• Hemin Substrate
• Hemin Standard (1 nmol)
Storage Conditions 4°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Free hemin results from the breakdown of hemin-containing proteins such as hemoglobin and myoglobin. It can be detected in various body fluids such as saliva, urine and csf under various pathological states. Free hemin exists in cells at a very minute concentrations (< 1µM ≈ 650 ng/ml) exerting regulatory functions such as repression of nonspecific δ-aminolevulinate synthase expression and induction of microsomal hemin oxygenase-1. Hemin can stimulate growth of oral bacteria involved in gingivitis and is an indicator of possible pathological conditions when found in the urine or feces. BioVision’s Hemin Assay Kit utilizes peroxidase activity in the presence of hemin to provide a simple, exquisitely sensitive assay which causes the conversion of a colorless probe to a strongly colored (λ = 570) compound. Trace amounts of Hemin can be quantitated in the 5-160 pg (10-250 fmol) range.


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Is this kit suitable for the measurement of free heme in plasma?
Free hemin exists in cells at a very minute concentrations (< 1µM ˜ 650 ng/ml) and this kit enable you to detect this in plasma samples.
Can we use this total liver lysate(stored at -20C or -80C) for estimating the Heme level, after estimating the protein and express the heme level in nmole/ug of liver protein?
You can prepare cell lysates using 10-100mg of mouse liver and adding 3-4 volumes of the assay buffer provided in the kit. The lysate can be stored at -80C but we recommend avoiding repeated freeze-thaw cycles. You can quantify the protein in the sample and add sample to each well based on amount of protein (final result can be expressed as amount of heme per ug lysate protein).
Can this kit quantify hemoglobin?
The hemin assay kit detects free heme. Hemoglobin has heme in coordinated form with protein chains of alpha and beta globin. Heme bound in this form will not produce color in this assay.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Is it possible to use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. It is recommended to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted.
Will the kit measure free hemin or bound hemin?
The kit measures free and it cannot measure bound hemin. Bound hemin will not generate any color during the reaction.
Flint & Stintzi. Cj1386, an Atypical Hemin-Binding Protein, Mediates Hemin Trafficking to KatA in Campylobacter jejuni. J. Bacteriol., Mar 2015; 197: 1002 - 1011.
Annika Flint et al., Cj1386 Is an Ankyrin-Containing Protein Involved in Heme Trafficking to Catalase in Campylobacter jejuni, J. Bacteriol., Jan 2012; 194: 334 - 345.
Zhiwei Huang, et. al. Sampangine Inhibits Heme Biosynthesis in both Yeast and Human. Eukaryot. Cell, Nov 2011; 10: 1536 - 1544.
For more citations of this product click here