Human MICB ELISA Kit (ab100593)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Equilibrate all reagents to room temperature (18-25°C) prior to use.	-Avoid foaming or bubbles when mixing or reconstituting components. -Ensure plates are properly sealed or covered during incubation steps. -Complete removal of all solutions and buffers during wash steps is necessary to minimize background.	-Unused well strips should be returned to the plate packet and stored at 4°C. -Don’t store the diluted HRP-Streptavidin solution for next day use.

Upstream tips
-Equilibrate all reagents to room temperature (18-25°C) prior to use.
Protocol tips
-Avoid foaming or bubbles when mixing or reconstituting components. -Ensure plates are properly sealed or covered during incubation steps. -Complete removal of all solutions and buffers during wash steps is necessary to minimize background.
Downstream tips
-Unused well strips should be returned to the plate packet and stored at 4°C. -Don’t store the diluted HRP-Streptavidin solution for next day use.

Publication protocol

Soluble MICB serum levels were measured in the sera from the HBV infected patient groups and in healthy controls using a commercially available MICB Human ELISA Kit (Abcam, Tokyo, Japan; Catalog No. ab100593) following the manufacturer’s instructions. Briefly, standard and sample were added into appropriate wells of coated ELISA plate and incubated over night at 4°C with gentle shaking. Plates were washed with wash solution and subsequently biotinylated MICB detection was added to each well and was incubated at room temperature with gentle shaking. After removing the solution and washing the plates, HRP-Streptavidin solution was added to each well and incubated at room temperature with gentle shaking. The plates were washed again and TMB One-Step substrate reagent was added to each well, incubated at room temperature in the dark with gentle shaking. Finally, stop solution was added to each well and plates were read at 450 nm immediately. The standard curve was plotted log-log graph paper based upon the mean of absorbance and the best fit straight line was drawn through the standard points. The concentrations of sMICB serum protein were calculated based upon the standard curve. The minimum detectable limit of sMICB serum proteins was 0.069 ng/ml.

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Manufacturer protocol

Download the product protocol from Abcam for Human MICB ELISA Kit (ab100593) below.

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