Ribose 5-Phosphate Isomerase A (RPIA) ELISA 试剂盒

• Pre-coated 96-Well Microplate
• Standard
• Standard Diluent Buffer
• Wash Buffer
• Detection Reagent A
• Detection Reagent B
• Diluent A
• Diluent B
• TMB Substrate
• Stop Solution
• Plate Sealer

• 37 °C incubator
• Multi and single channel pipettes and sterile pipette tips
• Squirt bottle or automated microplate washer
• 1.5 mL tubes
• Distilled water
• Absorbent filter papers
• 100 mL and 1 liter graduated cylinders
• Microplate reader (wavelength: 450 nm)
• ELISA Shaker

This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• 1) Standard: Prepare the standard with the recommended volume of Standard Diluent Buffer, to make the standard solution. Then use the Standard Diluent buffer to carry out serial dilutions of the standard solution, as instructed in the Protocol.

• 2) Wash Buffer: Dilute the concentrated Wash Buffer with distilled water, as instructed in the Protocol.

• 3) Detection Reagent Preparation: Calculate the total volume of working solution required. Dilute Detection Reagent A and Detection Reagent B with Diluent A and Diluent B, respectively, at 1:100.

This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• 1) Set standard, test samples and control wells.

• 2) Aliquot 50 μL of diluted standard into the standard wells.

• 3) Aliquot 50 μL of Standard Diluent buffer into the control (zero) well.

• 4) Aliquot 50 μL of diluted samples into the sample wells.

• 5) Immediately aliquot 50 μL of Detection Reagent A to each well. Incubate for 1 hr at 37 °C.

• 6) Wash 3 times.

• 7) Aliquot 100 μL of Detection Reagent B to each well. Incubate for 30 mins at 37 °C.

• 8) Wash 5 times.

• 9) Aliquot 90 μL of TMB Substrate to each well. Incubate for 10-20 mins at 37 °C.

• 10) Aliquot 50 μL of Stop Solution.

• 11) Measure the OD at 450 nm.

This assay is competitive, therefore there is an inverse correlation between RPIA concentration in the sample and the absorbance measured. Create a graph with the log of the standard concentration (y-axis) and average absorbance measured (x-axis). Apply a best fit trendline through the standard points. The RPIA concentration of the samples can be interpolated from the standard curve.