The purified recombinant gH/gL was dissolved in 10 mM PBS at pH 7.4. The 96-well microtiter plates were coated with 450 ng of gH/gL protein per well at 37°C for 2 h. After incubation, unoccupied sites were blocked with 3% BSA in PBS. The plates were washed five times with PBS containing 0.1% Tween-20 (PBS-T). After washing, serum samples (1:100 in PBST containing 3% BSA) were added and incubated for 1 h at 37°C. After five washes, HRP-labelled goat anti-human IgA antibodies (1:6000; Boster corporation, Wuhan, China) for gH/gL in PBS-T were added. The plates were incubated at 37°C for 30 min and washed before adding a tetramethylbenzidine reagent (Sigma-Aldrich, St Louis, MO, USA) for 10 min at 37°C. The reaction was stopped with 2 M H2SO4 and the optical density (OD) at 450 nm was determined using an ELISA reader.
Tetramethylbenzidine reagent
Manufactured by Merck Group
Sourced in United States
Tetramethylbenzidine reagent is a colorimetric substrate used in various analytical and diagnostic assays. It is a chemical compound that undergoes an enzymatic reaction, resulting in a color change that can be detected and measured. The core function of this reagent is to serve as a sensitive indicator in these analytical procedures.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using tetramethylbenzidine reagent
1
ELISA for gH/gL-Specific Antibodies
2
Quantifying Lactic Acid in Multiple Myeloma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Lactic acid immunoassay kits (Y4324; Yearthbio) were used to determine the lactic acid levels in MM cells, according to the manufacturer's instructions. Briefly, the lactic acid antibody (1:100; included in the kit) was incubated with MM cells overnight at 4°C. Following this, the MM cells were incubated with horseradish peroxidase-labeled anti-rabbit antibody (1:5,000; included in the kit) for 30 min at room temperature. Wells were then developed with tetramethyl benzidine reagent (Sigma-Aldrich; Merck KGaA) in the dark and the absorbance was measured at 450 nm.
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