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Home / Assay Kits

MicroMolar Histidine Assay Kit

SKU : BHE16300252

This product includes 750 ul 10 x C53 Dye and 50 ul of 10 mM Histidine. It is for measurement of 200 samples using 96-well plates. Cuvettes may also be used for measurements.

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Mfr.No.	HIS200
Description	Histidine is an important molecule in biology and pharmaceutical science. Metabolic block of histidine results in increased concentrations of histidine in blood, urine, and cerebrospinal fluid. Disorders of histidine metabolism is related to diseases such as histidinemia and urocanic aciduria. Histidine is a common ingredient of pharmaceutical products. The MicroMolar Histidine Assay Kit is designed for measurement of micromolar concentrations of histidine. The assay is based on increase of fluorescence at 535 nm of the dye C53 in the presence of histidine. The assay kit can be used for measurements histidine concentrations in biological samples, biochemical reactions, pharmaceutical products and environmental water samples. The assay is compatible with HEPES buffer, low concentrations of nonionic detergent (<0.01%), MgCl2 (< 5 mM), CaCl2 (<5 mM), EDTA (< 1 mM) and phosphate (< 1 mM). It is not compatible with thiol compounds such as DTT. For measurement of other amino acids, see the information of Amino Acid Assay Kit (Catalog Number AAK1000). The MicroMolar Histidine Assay Kit (catalog number HIS200) includes 750 µl 10 x C53 Dye and 50 µl of 10 mM Histidine. It is for measurement of 200 samples using 96-well plates. Cuvettes may also be used for measurements.
Concentration measurement	Biomolecules

Product Manual	Datasheet
Protocols Description	ASSAY PROTOCOL The following assay protocol is based on using a 96-well plate for the measurement. The sample volume is 150 µl and the final assay volume is 188 µl. For 384-well plate assays, the sample volume is 60 µl and the final assay volume is 75 µl. For assays using cuvette, the sample volume is 800 µl and the final assay volume is 1000 µl. STANDARD CURVE 1. Sample preparation: Prepare 150 µl of histidine solutions in the wells of a black 96-well plate with a two-fold serial dilution from 0.050 mM to zero in a 10 mM HEPES, pH 7.4 buffer. For 10 samples, dilute 40 µl of the 10 x C53 dye 10-fold with water to make 400 µl of 1 x C53 dye. 2. Detection: Mix 37.5 µl of 1 x dye C53 with 150 µl of the histidine solutions for 5 min and read the fluorescence at 535 nm (excitation at 485 nm). 3. Data Analysis: Plot the fluorescence intensity Fc and the histidine concentration [Histidine] to generate the linear standard curve. Fc = a [Histidine] + b Where the Fc values are from experimental data, the a and b values are from the linear fitting between the Fc values and the histidine concentrations. UNKNOWN SAMPLES Follow the same procedure to measure the fluorescence intensity Fc values from the unknown samples. Calculate the histidine concentrations in the unknown samples using the Fc values from the unknown samples and the a and b values from the standard curve. [Histidine] = (Fc – b) / a

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SKU	Supplier No.	Size	Your price	Quantity
BHE16300252	HIS200	200 assays	$302.17		Add to Cart

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