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MicroGram Lipid Assay Kit

BHE16300268

This product includes 0.1 ml of 1000 x LIP 10 Dye. It is for measurement of 1000 samples using 96-well plates. Cuvettes may also be used for measurements.

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Product Manual Datasheet
Protocols Description ASSAY PROTOCOL
The following assay protocol is based on using a 96-well plate for the measurement.
 
Protocol 1
Most lipids can be measured using protocol1.
Add 100 µl of 0.1 M NaCl into 12 wells of a black 96-well plate. Mix100 µl of 1 mg/ml lipid in ethanol or methanol with the first well containing 100 µl of 0.1 M NaCl. Perform a two-fold serial dilution in the other wells containing 100 µl of 0.1 M NaCl and leave the last well for zero control. For each 20 samples, dilute 2 µl of 1000 x LIP10 Dye with 2 ml of water to make the 1 x LIP10 Dye. Mix 100 µl of LIP10 Dye with 100 µl of the lipid solutions and incubate the mixture for 5 min. Read the fluorescence at 465 nm (excitation at 360 nm).
 
Protocol 2
Some lipids should be measured using protocol 2 due to its insolubility in an aqueous solution. Add 100 µl of methanol into 12 wells of a black 96-well plate. Mix100 µl of 1 mg/ml lipid in ethanol or methanol with the first well containing methanol. Perform a two-fold serial dilution in the other wells containing 100 µl of methanol and leave the last well for zero control. For each 20 samples, dilute 2 µl of 1000 x LIP10 Dye with 4 ml of water to make the 0.5 x LIP10 Dye. Mix 200 µl of the 0.5 x LIP10 Dye with 100 µl of the lipid solutions and incubate the mixture for 5 min. Read the fluorescence at 465 nm (excitation at 360 nm).
 
Data Analysis: Plot the fluorescence intensity Fc and the lipid concentration [Lipid] to generate the linear standard curve.
 
                                                                    Fc = a [Lipid] + 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 lipid concentrations.
 
UNKNOWN SAMPLES
Dissolve the lipid sample in ethanol or methanol. Follow the same procedure to measure the fluorescence intensity Fc values from the unknown samples. Calculate the lipid concentrations in the unknown samples using the Fc values from the unknown samples and the a and b values from the standard curve.
 
                                                                   [Lipid] = (Fc – b) / a
SKU Supplier No. Size Your price Quantity
BHE16300268 LIP1000 1000 assays $329.13
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