MicroMolar DTT Assay kit

Mfr.No.	DTT200
Description	DTT (Dithiothreitol) is a common reducing agent in biochemistry. Removal of DTT is required for Cys-based protein labeling and for disulfide bond formation in proteins. The MicroMolar DTT Assay Kit (Catalog number DTT200) is designed for measurement of micromolar concentrations of DTT. The assay is based on increase of fluorescence at 535 nm of the dye C55 in the presence of DTT. The assay kit can be used for measurements DTT concentrations in biological samples, biochemical reactions and environmental water samples. The assay is compatible with HEPES buffer, low concentrations of non-ionic detergent (<0.01%), MgCl2 (< 5 mM), CaCl2 (<5 mM), Tris-HCl (<10 mM), EDTA (< 1mM) and phosphate (< 1 mM). It is not compatible with thiol compounds such as 2- mercaptoethanol or cysteine. The MicroMolar DTT Assay Kit (catalog number DTT200) includes 5 ml of Dye C55. It is for measurement of 200 samples using 96-well plates. Cuvettes may also be used for measurements.
Concentration measurement	Salt and buffer components

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 100 µl and the final assay volume is 125 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 100 µl of DTT solutions in the wells of a black 96-well plate with a two-fold serial dilution from 0.1 mM to zero in water or a 10 mM HEPES, pH 7.4 buffer. 2. Detection: Mix 25 µl of Dye C55 with 100 µl of the DTT solutions for 10 min and read the fluorescence at 535 nm (excitation at 485 nm). 3. Data Analysis: Plot the fluorescence intensity Fc and the DTT concentration [DTT] to generate the linear standard curve. Fc = a [DTT] + 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 DTT concentrations. UNKNOWN SAMPLES Follow the same procedure to measure the fluorescence intensity Fc values from the unknown samples. Calculate the DTT concentrations in the unknown samples using the Fc values from the unknown samples and the a and b values from the standard curve. [DTT] = (Fc – b) / a

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SKU	Supplier No.	Size	Your price	Quantity
BHE16300283	DTT200	200 assays	$320.18		Add to Cart