Gentaur Alanine Aminotransferase
Alanine aminotransferase 1 mediates the control of a large number of enzymes, ion channels, aquaporins and other proteins throug calcium-binding. Among the enzymes to be stimulated by the calmodulin-calcium complex are a number of protein kinases and phosphatases.
Together with CCP110 and centrin, is involved in a genetic pathway that regulates the centrosome cycle and progression through cytokinesis. BioVision’s GPT ELISA kit is a sandwich ELISA assay for the quantitative measurement of rat GPT in serum, plasma and cell culture supernatants. The density of color is proportional to the amount of rat GPT captured from the samples.
- Prepare all reagents, samples and standards as instructed in section VII.
- Wash plate 2 times with 1X Wash Solution before adding standard, sample and control wells.
- Add 100 μl of each standards or samples into appropriate wells. Cover well and incubate for 1.5 hours at 37ºC.
- Remove the cover and discard the plate content without washing or letting the wells completely dry.
- Discard the solution and wash 3 times with 1X Wash Solution. Wash by filling each well with Wash Buffer (350 μl) using a multi-channel
- pipette or autowasher. Let it soak for 1-2 minutes, and then remove all residual wash-liquid from the wells by aspiration. After the last
- wash, remove any remaining Wash Buffer by aspirating or decanting. Clap the plate on absorbent filter papers or other absorbent
- Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37ºC for 30 min.
- add 90 μl of TMB substrate into each well, cover the plate and incubate at 37 ºC in dark within 15-30 min. The shades of blue should be seen in the first 3-4 wells by the end of incubation
- Add 50 μl of Stop Solution to each well. Read result at 450 nm within 20 minutes.