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EL-C1600N100013-B
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2015/03/31 09:38:55 The other party has successfully received the offline file "Human Staphylococcal Protein A (SPA) Manual.pdf" (148.91KB).
The following is a human staphylococcal protein A (SPA) ELISA kit for our customers to organize the experimental steps:
Human staphylococcal protein A (SPA) ELISA kit experimental materials and reagent preparation: 1. Instruments and materials: microplate reader (preheating for 30 minutes before use), micro-adjuster, pipette, distilled or deionized water, filter paper . 2. Preparation of the lotion: Prepare the lotion at a ratio of 1:30. Human Staphylococcal Protein A (SPA) ELISA Kit Procedure: 1) Remove the kit and leave it at room temperature (20-25 ° C) for 30 minutes. 2) Grouping: Take out the 96-well plate, determine the number of slats required according to the number of samples to be tested plus the number of standards, and continue to refrigerate the remaining slats. Set standard group (6 concentrations), blank holes, and sample groups to be tested. 3) Dilution of standard products: Prepare 6 small test tubes, and edit the numbers one by one. Add 100 ul of standard dilution solution to each small test tube, then take 100 ul of the original concentration standard and add it to a well-tested test tube. Mix well; then take 100 ul in the test tube and add to the second test tube, mix well; then take 100 ul in the test tube and add to the third test tube, mix well; then take 100 ul in the test tube and add the fourth In the test tube, mix thoroughly; then add 100 ul to the fifth tube in the test tube and mix well; then take 100 ul in the tube and discard. The sixth tube was used as the No. 0 standard. Note: In order to reduce the experimental error and ensure the accuracy, it is recommended to set the double hole. 4) Loading: Add 50 ul of standard solution to the blank microwell according to the order of the standard; add 50 ul of distilled water to the blank control well; add 40 ul of the sample to the remaining microwell and then add 10 ul of the biotin-labeled antibody. Add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, and shake gently to mix. 5) Incubation: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes. 6) Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing solution, let stand for 30 seconds, and repeat it 5 times and pat dry. 7) Add the enzyme standard solution: 50 ul of the enzyme standard solution (except the blank control hole) is added to each well of the standard group and the sample group to be tested. 8) Incubation: The enzyme-labeled plate was sealed with a sealing paper, and then placed in a wet box and incubated at 37 ° C for 1 hour at a constant temperature. 9) Washing the plate: Fill each well with the diluted washing solution, let stand for 15-30 s, thoroughly clean the plate for 5 times, and pat dry thoroughly with absorbent paper. 10) Color development: 50 ul of the developer A liquid was added to each well, and then 50 ul of the developer B solution was added. 11) Termination: 10-15 minutes in the dark at 25-37 ° C, add 50 ul of stop solution. 12) Plate reading: The OD value of each well was read at a wavelength of 450 nm. Note: The liquid and finger marks remaining at the bottom of the board must be wiped out when reading the board. The reading time is controlled within 30 minutes after the termination of the reaction, so as not to affect the accuracy.
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