Prolactin Rat ELISA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Reagents with different lot numbers should not be mixed. -All reagents need to be brought to room temperature prior to use.	-Substrate Solution should remain colourless until added to the plate. Keep Substrate Solution protected from light . -Stop Solution should remain colourless until added to the plate. The colour developed in the wells will turn from blue to yellow immediately after the addition of the Stop Solution. Wells that are green in colour indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Upstream tips
-Reagents with different lot numbers should not be mixed. -All reagents need to be brought to room temperature prior to use.
Protocol tips
-Substrate Solution should remain colourless until added to the plate. Keep Substrate Solution protected from light . -Stop Solution should remain colourless until added to the plate. The colour developed in the wells will turn from blue to yellow immediately after the addition of the Stop Solution. Wells that are green in colour indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Publication protocol

A pipetted 25 μl of each sample into the wells was prepared and added 50 μl of rat prolactin sample buffer to every well. Shake for 2 h at room temperature (22 ± 1 °C), rinsed 4 times with 300 μl buffered wash solution. A 200 μl was added of enzyme-labeled anti-rat prolactin antibody to all wells and shaken for 1 h, rinsed again for 4 times with 300 μl buffered wash solution. A 200 μl of liquid TMB/substrate solution was added to all wells. This was then incubated standing for 30 min in the dark. Then 50 μl of stop solution was added to each well, mixed carefully and measured at 450 nm

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Manufacturer protocol

Download the product protocol from BioVendor for Prolactin Rat ELISA below.

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