Detection kit for quantitatively detecting heart-type fatty acid binding protein by ELISA method
A fatty acid combination and detection kit technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of poor uniformity, low stability, and long detection time of kits, and achieve low price and good uniformity , the effect of short detection time
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Embodiment 1
[0052] The composition of embodiment one kit:
[0053]The embodiment of the present invention provides a detection kit for the quantitative detection of heart-type fatty acid binding protein by ELISA, including a microtiter plate coated with an anti-H-FABP monoclonal antibody, and an anti-H-FABP polyclonal antibody for HRP-labeled detection , standard, standard diluent, sample diluent, concentrated washing solution, chromogenic solution A, chromogenic solution B, stop solution, detection antibody diluent.
[0054] The components and preparation of the materials contained in the kit are as follows:
[0055] The preparation method of the enzyme label plate coated with anti-H-FABP monoclonal antibody comprising the following steps:
[0056] 1) Dissolving the coated anti-H-FABP monoclonal antibody in the coating buffer until the concentration of the coating antibody is 2ug / mL to obtain a mixed solution;
[0057] 2) Add the mixed solution obtained in step 1) into the wells of t...
Embodiment 2
[0079] Embodiment two: the step that detects with kit
[0080] (1): Preparation of standard curve
[0081] Prepare the following solutions before use:
[0082] (1) Dilute the HRP-labeled anti-H-FABP polyclonal antibody for detection with the detection antibody diluent, so that the ratio of solute (anti-H-FABP polyclonal antibody for detection) to solvent (detection antibody diluent) is 1:5000;
[0083] (2) Dissolve the standard with the standard diluent so that the concentration of the standard is 0pg / mL, 5pg / mL, 15pg / mL, 40pg / mL, 130pg / mL, 300pg / mL.
[0084] The detection steps are as follows:
[0085] 1. Dosing: Dilute the concentrated washing solution (20×) with distilled or deionized water 1:20 for later use (30mL washing solution is diluted to 600mL, 50mL washing solution is diluted to 1000mL);
[0086] 2. Numbering: Number the corresponding microwells of the sample in sequence;
[0087] 3. Adding samples: Add 50 μL of the calibrator solution to the corresponding we...
Embodiment 3
[0101] Embodiment three, the performance of kit
[0102] The kit to be tested is: prepare three batches of kits (batch one, batch two and batch three)
[0103] (1) Minimum detection limit
[0104] 1. Take the microwell plate coated with H-FABP, add 50 μL standard substance to each well and shake gently to mix. Incubate at 37°C for 30 minutes. Wash the plate 5 times, add 100 μL of enzyme conjugate to each well, and incubate at 37°C for 30 minutes. For each kit to be tested, 20 duplicate holes were set for the S0 solution, and double holes were added for other calibrators.
[0105] 2. After completing step 1, fully wash 5 times with washing liquid, and buckle dry.
[0106] 3. After completing step 2, add 50 μL of chromogenic solution A and 50 μL of chromogenic solution B to each well, and develop color at 37°C in the dark for 15 minutes. Add 50 μL of stop solution to each well, and use 450 / 630 nm dual wavelength detection on a microplate reader. The absorbance values of ...
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