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ELISA detection kit and preparation method thereof

A detection kit and enzyme-linked technology, applied in the field of biomedicine

Active Publication Date: 2012-05-02
北京健平九星生物医药科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Houck et al. (J Biol. Chem. 1992, 267:26031) reported an ELISA method for detecting VEGF based on colorimetry, but its detection sensitivity was only ng / ml

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The kit of the present invention can be exemplified by the following substances:

[0023] (1) VEGF receptor coated enzyme-linked plate;

[0024] (2) Positive control (VEGF protein freeze-dried powder) 1 tube;

[0025] (3) Negative control (animal serum freeze-dried powder) 1 tube;

[0026] (4) Sample diluent: 0.05% Tween-20, 1% bovine serum albumin, 10mM disodium edetate, 0.05% thimerosal, 0.4M NaCl, pH6.5, 1 bottle;

[0027] (5) Enzyme conjugate: horseradish peroxidase-labeled vascular endothelial growth factor monoclonal antibody, 1 vial;

[0028] (6) The concentrated washing solution is: 137mM NaCl, 2.7 mM KCl, 10mM NaCl 2 HPO 4 , 2 mM KH 2 PO 4 , 0.05% Tween-20, Ph7.4, 1 bottle;

[0029] (7) Chromogenic agents A and B, of which the liquid level of A is H 2 o 2 solution, solution B is 3, 3 ＇ ,5 ,5 ＇ -Tetramethylbenzidine solution, 1 bottle each;

[0030] (8) The stop solution is 2MH 2 SO 4 , 1 bottle;

[0031] (9) 2 sheets of sealing tape.

[0032] ...

Embodiment 2

[0090] Embodiment 2: Kit use method

[0091] 1. Take out the required strips from the airtight bag equilibrated to room temperature, and seal the other strips and put them back at 4°C.

[0092] 2. Leave one well as a blank well, and add 100 μl (or 2 drops) of sample diluent to the other wells.

[0093] 3. In addition to the blank well, add 100 μl each of negative control well 1, positive control 2 wells, and serum samples to the corresponding wells, and mix well by flicking. Seal the reaction well with sealing tape and keep at 37°C for 60 minutes.

[0094] 4. Wash the plate 3 times: (1) Automatic plate washer: Shake off the liquid in the well, the washing liquid required to be injected is 350μl, the interval between injection and aspiration is 15-30 seconds, and the plate is washed 3 times. (2) Wash the plate manually: Shake off the liquid in the wells, add 350 μl of washing solution to each well, let it stand for 30 seconds, shake off the liquid, pat dry on thick absorbent ...

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PUM

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Abstract

The invention provides an ELISA detection kit used for detecting human vascular endothelial growth factors (VEGF). In the detection kit, there is a VEGF acceptor coated enzyme-linked reaction plate, a enzyme conjugate prepared from horseradish peroxidase (HRP) labeled antibodies and a protective agent, lyophilized powder of VEGF protein used as positive contrast, lyophilized powder of protein used as negative contrast, sample diluents, PBST concentrated lotion, a chromogenic substrate prepared from 3,3',5,5'-tetramethyl benzidine, 2MH2SO4 used as a stopping solution and shrouding glue paper. The kit can be used for diagnosis and prognosis of cancers, angiopathy, diabetic retinopathy and rheumatoid arthritis and has the advantages of accuracy, specificity, sensitivity, stability, convenience and the like.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to an enzyme-linked detection kit for detecting human vascular endothelial cell growth factor (VEGF). [0002] The present invention also relates to the preparation method of the above kit. Background technique [0003] Tumor development and metastasis are multi-stage, complex and highly selective processes involving the intricate relationship between tumor cells and hosts, and the mechanism is still unclear. In recent years, research on the relationship between new angiogenesis and tumor development, metastasis, and prognosis has progressed rapidly, proving that new angiogenesis is a necessary condition for tumorigenesis, metastasis, and the growth of metastatic foci, and vascular endothelial growth factor (VEGF) is the regulator of neovascularization. The most important cytokine produced. Studies have shown that vascular endothelial growth factor (VEGF) has an irreplaceable ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558
Inventor 邹检平
Owner 北京健平九星生物医药科技有限公司
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