Ctenopharyngodon idellus interleukin-1beta (IL-1beta) enzyme-linked immunosorbent assay (ELISA) kit

An enzyme-linked immunosorbent assay and interleukin technology, applied in the biological field, can solve the problems of lack of establishment, large differences in molecular structure, and inability to identify, and achieve the effect of low cost

Inactive Publication Date: 2014-12-24
UNIV OF ELECTRONICS SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the method for detecting the secretion of interleukin 1β from grass carp has not been established, mainly because the molecular structure of interleukin 1β from grass carp is quite different from that of mammals and amphibians, and the interleukin 1β antibody of other species cannot be used. to identify grass carp interleukin-1β

Method used

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  • Ctenopharyngodon idellus interleukin-1beta (IL-1beta) enzyme-linked immunosorbent assay (ELISA) kit
  • Ctenopharyngodon idellus interleukin-1beta (IL-1beta) enzyme-linked immunosorbent assay (ELISA) kit
  • Ctenopharyngodon idellus interleukin-1beta (IL-1beta) enzyme-linked immunosorbent assay (ELISA) kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The composition of embodiment 1 kit of the present invention and its using method

[0028] 1. The composition of kit of the present invention

[0029] (1) 96-well detachable microplate;

[0030] (2) grass carp interleukin-1β polyclonal antibody concentrate labeled with horseradish peroxidase;

[0031] (3) Sample diluent: washing buffer containing 0.2% bovine serum albumin (BSA), containing KH in every liter of dilution buffer 2 PO 4 0.2g, Na 2 HPO 4 2.9g, NaCl8.0g, BSA2.0g, Tween-200.5ml, PH7.4;

[0032] (4) Concentrated washing solution: 0.2M phosphate buffer containing 0.5% Tween-20, containing KH in every liter of concentrated washing solution 2 PO 4 2.0g, Na 2 HPO 4 29.0g, NaCl80.0g, Tween-205.0ml, PH7.4;

[0033] (5) Concentrated coating solution: 0.5M carbonate buffer solution, each liter of concentrated coating solution contains NaCO 3 15.9g, NaHCO 3 29.3g, pH9.6;

[0034] (6) Chromogenic substrate: soluble one-component TMB substrate solution;

[0...

Embodiment 2

[0058] Embodiment 2 The verification of the reliability of the kit of the present invention

[0059] 1. The linear range detection of the kit of the present invention

[0060] (1) Method: The prepared recombinant grass carp interleukin 1β standard series solutions (0, 0.4, 2, 10, 50, 100 ng / ml) were tested according to the above ELISA detection method, and the inhibition rate of each concentration was calculated. Taking the inhibition rate as the vertical axis and the logarithm of the recombinant grass carp interleukin-1β solution concentration as the horizontal axis, draw the standard inhibition curve.

[0061] (2) Results: see Table 2 for competition ELISA results and related parameters, see the standard inhibition curve figure 1 , it can be seen that within the range of 0.4-100ng / m1, the kit of the present invention has good linearity.

[0062] Table 2 Related parameters of competitive inhibition ELISA results

[0063]

[0064] (3) Conclusion: the kit of the present i...

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Abstract

The invention belongs to the biotechnology field and discloses a ctenopharyngodon idellus interleukin-1beta (IL-1beta) competitive inhibiting enzyme-linked immunosorbent assay (ELISA) kit. The kit comprises the following components: (1) an ELISA plate, (2) a horse radish peroxidase marked ctenopharyngodon idellus IL-1beta polyclonal antibody concentrated solution, (3) a sample diluent, (4) washing concentrate, (5) concentrated coating buffer, (6) a chromogenic substrate, (7) stop buffer and (8) standard ctenopharyngodon idellus IL-1beta. The kit has the characteristics of simplicity in operation, good repeatability, accuracy and specificity and wide linear detection range.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a grass carp interleukin-1β competitive inhibition ELISA detection kit. Background technique [0002] Interleukin-1β (interleukin-1β, IL-1β) is an important inflammatory mediator and immune factor, which plays a wide range of regulatory roles in the body's immune response, such as activating the activity of T cells, B cells and natural killer cells , stimulate macrophages to secrete cytokines and prostaglandins and other inflammatory mediators, and also act as a signal transmission between the nervous system and immune system to monitor physiological disorders and pathogen invasion. As a potential inflammatory mediator and co-stimulatory signal molecule, interleukin-1β can promote the synthesis of prostaglandins (PG), induce fever, increase the phagocytic activity of macrophages, and regulate the expression of other cytokines. [0003] Grass carp (Ctenopharyngodon idellu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6869
Inventor 周红杨潇
Owner UNIV OF ELECTRONICS SCI & TECH OF CHINA
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