• SENSITIVITY : 3000 NG/WELL.
• RANGE : 3000 NG/WELL – 25000 NG/WELL.
• ASSAY TYPE: SANDWICH (QUANTITATIVE).
• SAMPLE TYPE : SERUM, PLASMA AND OTHER BIOLOGICAL FLUIDS.

This procedure is provided for reference only. The product manual may differ slightly. The product must be used as described in the product manual included and delivered with the product.
Equilibrate kit components and samples at room temperature (18 – 25 °C) prior to use. It is recommended to draw a standard curve for each test.

1. Place the wells of the standard, test sample and control (zero) respectively on the pre-coated plate and record their positions. It is recommended to measure each standard and each sample at least twice.
2. Add 100 µl of each standard, control and sample to the appropriate wells. Seal the plate with a lid and incubate for 1 hour at 37°C.
3. Remove the lid and discard the liquid.
4. Add 100 µL of detection reagent A working solution to each well. Close plate with lid and incubate for 1 hour at 37°C.
5. Remove the lid and discard the solution. Wash the plate three times with 1X Wash Buffer.
6. Add 100 µL of Detection Reagent B working solution to each well, seal and incubate at 37°C for 30 minutes.
7. Discard the solution and wash the plate 5 times with Wash Buffer as described in the previous step.
8. Aliquot 90 µl of TMB substrate into each well. Close the plate with a lid and incubate at 37°C for 10-20 minutes. Avoid exposure to light. The incubation time is given as a guide only, the optimal time should be determined by the end user. Do not exceed 30 minutes.
9. Add 50 µl of Stop Solution to each well. Read immediately at 450 nm.

The Ras ELISA kit uses the competitive enzyme immunoassay technique. It is based on the use of a monoclonal antibody to Ras protein and a Ras-HRP protein conjugate. The test matrix and buffer are incubated with the Ras-HRP protein conjugate in a pre-reusable container for one hour. After the incubation period, samples are decanted and washed five times. Then the wells are incubated with a substrate for the HRP enzyme. The product of the enzyme-substrate reaction forms a blue complement. Finally, a water sample is added. Finally, a parachute solution is added to stop the reaction. The intensity of the colour is measured spectrophotometrically at 450 nm in a microplate reader. Microplate reader.

The colour intensity is inversely proportional to the concentration of the Ras protein. Ras ligands and Ras-HRP conjugate are mixed with Ras anti-protein. The intensity of the colour is inversely proportional to the concentration of the Ras protein. Since the number of sites is limited, the more sites are occupied by the machine Ras protein, the fewer sites are available for binding to the conjugated Ras-HRP protein. A linear curve relates the colour intensity (D.O.) to the concentration of the parameters. The concentration of Ras protein in the sample.

The concentration of Ras protein in each sample is calculated from this curve. Principle of the test This kit is based on sandwich immunoassay technology. An antibody is pre-coated on a 96-well plate. Standards, test samples and biotin conjugate reagent are added to the wells and incubated. HRP conjugate reagent is then added and the entire plate is incubated. Unbound conjugates are removed with Wash Buffer in each step. TMB substrate is used to quantify the HRP enzyme reaction.

After addition of the TMB substrate, only wells with sufficient RRAS2 produce a blue coloured product, which turns yellow after addition of the acid stop solution. The intensity of the yellow colour is proportional to the amount of RRAS2 bound to the plate. The optical density (OD) is measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of RRAS2 can be calculated.