R & D Systems offers a complete range of complete ELISA kits. To ensure high quality and repeatability, all kits have undergone extremely strict quality inspections. To detect intracellular and extracellular proteins, the kit uses a variety of detection methods, including colorimetry, fluorescence and chemiluminescence. Specification: 48 times / 96 times Price: 1600/2600 96 times kit composition: Detection range: see information This kit is used to determine the content of placental alkaline phosphatase in human serum, plasma and related liquid samples. If you want to know whether the price of the human placental alkaline phosphatase ELISA kit is discounted, special price, or kit related information, please consult our sales staff:
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1 30 times concentrated washing solution 20ml × 1 bottle 7 stop solution 6ml × 1 bottle
2 Enzyme label reagent 6ml × 1 bottle 8 standard (pg / ml) 0.5ml × 1 bottle
3 Enzyme label coating plate 12 wells × 8 strips 9 Standard diluent 1.5ml × 1 bottle
4 Sample diluent 6ml × 1 bottle 10 Instructions 1 copy
5 Developer A solution 6ml × 1 bottle 11 sealing film 2 sheets
6 Developer B liquid 6ml × 1 / bottle 12 sealed bag 1
Composition of 48 kits:
1 20 times concentrated washing solution 20ml × 1 bottle 7 stop solution 3ml × 1 bottle
2 Enzyme label reagent 3ml × 1 bottle 8 standard (pg / ml) 0.5ml × 1 bottle
3 Enzyme label coating plate 12 well × 4 strips 9 Standard dilution 1.5ml × 1 bottle
4 Sample diluent 3ml × 1 bottle 10 Instructions 1 copy
5 Developer A solution 3ml × 1 bottle 11 sealing film 2 sheets
6 Developer B solution 3ml × 1 / bottle 12 sealed bag 1 special case: human placental alkaline phosphatase ELISA kit English name: human placental alkaline phosphatase (PLAP) elisa kit
Experimental principle The kit uses the double antibody sandwich method to determine the level of human placental alkaline phosphatase in the specimen. Microporous plates were coated with purified human placental alkaline phosphatase antibody to prepare solid-phase antibodies. Placental alkaline phosphatase was added to the monoclonal antibody-coated microwells in turn, followed by HRP-labeled placental alkaline phosphatase antibody The combination forms an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid. The color depth is positively correlated with the placental alkaline phosphatase in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the concentration of human placental alkaline phosphatase in the sample was calculated by a standard curve.
Steps
1. Dilution of standard products: This kit provides one original standard product. The user can perform dilution in a small test tube according to the following chart.
8 ng / L No. 5 standard 150μl primary standard added 150μl standard dilution
4 ng / L No. 4 standard 150μl No. 5 standard added 150μl standard dilution
2 ng / L No. 3 standard 150μl No. 4 standard added 150μl standard dilution
1 ng / L No. 2 standard 150μl No. 3 standard added 150μl standard dilution
0.5 ng / L No. 1 standard 150 μl No. 2 standard added 150 μl standard dilution
2. Sample addition: set up blank wells (without adding samples and enzyme-labeled reagents in the blank control wells, the rest of the operations are the same), standard wells, and sample wells to be tested. Accurately add 50μl of the standard on the enzyme-coated plate, add 40μl of sample diluent to the sample well, and then add 10μl of the sample to be tested (the final dilution of the sample is 5 times). Add the sample and add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, shake gently to mix.
3. Incubation: seal the plate with a sealing film and incubate at 37 ° C for 30 minutes.
4. Mixing solution: dilute 20 times concentrated washing liquid with distilled water 20 times and reserve
5. Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with the washing solution, let it stand for 30 seconds and then discard, repeat this 5 times, pat dry.
6. Add enzyme: add 50μl of enzyme label reagent to each well, except blank well.
7. Incubation: The operation is the same as 3.
8. Washing: The operation is the same as 5.
9. Color development: Add 50μl of developer A to each well, and then add 50μl of developer B, mix gently, and develop at 37 ° C in the dark for 15 minutes.
10. Termination: Add 50μl of stop solution to each well to stop the reaction (at this time, the blue color turns to yellow).
11. Determination: Measure the absorbance (OD value) of each well in sequence with the blank air conditioner at zero and 450 nm wavelength. The measurement should be carried out within 15 minutes after adding the stop solution.
Storage conditions and validity period
1. Kit storage: 2-8 ° C.
2. Validity period: 6 months, transportation conditions, low temperature transportation, dry ice or ice pack low temperature transportation.
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Manual of human placental alkaline phosphatase (PLAP)
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