SRF 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

The stability of the kit is determined by the rate of activity loss. The loss rate is less than 5 % within the expiration date under appropriate storage conditions. To minimize performance fluctuations, operation procedures and lab conditions should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same user throughout.

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.

• Equilibrate the kit components and samples to room temperature (18 - 25 °C) before use. It is recommended to plot a standard curve for each test.

• 1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample at least in duplicate.

• 2. Add 100 μL of each standard, control and sample into the appropriate wells. Seal the plate with a cover and incubate for 1 h at 37 °C.

• 3. Remove the cover and discard the liquid.

• 4. Add 100 μL of the detection Reagent A working solution to each well. Seal the plate with a cover and incubate for 1 h at 37 °C.

• 5. Remove the cover and discard the solution. Wash the plate 3 times with 1X Wash Buffer.

• 6. Add 100 μL of Detection Reagent B working solution into each well, seal and incubate at 37 °C for 30 min.

• 7. Discard the solution and wash the plate 5 times with wash buffer as explained in previous step.

• 8. Aliquot 90 μL of TMB Substrate into each well. Seal the plate with a cover and incubate at 37 °C for 10-20 min. Avoid exposure to light. The incubation time is for reference use only, the optimal time should be determined by end user. Do not exceed 30 min.

• 9. Add 50 μL of Stop Solution to each well. Read at 450 nm immediately.

For calculation, average the O.D.450 duplicate readings for each reference standard and each sample and substract the average control (zero) O.D.450 reading. The standard curve can be plotted as the relative O.D.450 of each reference standard solution (Y) vs. the respective concentration of each standard solution (X). The SRF concentration of the samples can be interpolated from the standard curve.