Animal disease diagnostic Monkey malaria antibody ELISA detection kit

Simian malaria antibody ELISA kit

1.Principle

Specific genetic engineering antigen is coated on microtiter plate. After bounding with antibodies in the sample, enzyme conjugate is added, reacts with the antibody-antigen complex and generates a new substrate, unreacted materials are washed away. The addition of TMB solution leads to a color change, and anti-SM antibodies in the serum are detected.

2.Components

1) Antigen coated microwell........................................................ 1 piece(96 well)

2) Substrate A.............................................................................. 1 bottle(7 mL)

3) Substrate B............................................................................. 1bottle(7 mL)

4) Enzyme conjugate.................................................................. 1bottle(12 mL)

5) Sample diluent solution.......................................................... 1bottle(12 mL)

6) Stop solution........................................................................... 1 bottle(7 mL)

7) Positive control serum.............................................................. 1bottle(1 mL)

8) Negative control serum............................................................. 1 bottle(1 mL)

9) Cover membrance.............................................................................. 1piece

10) 20x washing buffer............................................................... 1bottle(50 mL)

11) Manual...................................................................................................... 1

12) Outo-bag Desiccant............................................................................ 1set

3.ELISA procedure

1) Take out all reagents to room temperture(18-25℃) for at least 30 min, and dilute 20xwashing buffer with deionized water to 1000 mL,shake properly for use.

2) Take out microplate from bag ,set blank control well ,only add sample diluent ,set 2 wells for negative control and add 100 μL negative control serum, set 2 wells for positive control and add 100 μL positive control serum; other wells ,add 100 μL sample diluent,first, add 10 μL test serum, shake properly and generate a blue-green color, put microplate in vortex to mix for 5-10 s, seal the microwell withcover membrance,resealed unsed microwell.

3) Incubate microplate at 37 ℃ for 30 min, pour liquid out of microwell , wash 5 times with washing buffer, place it for 20-30 s every time, flap to dry on absorbent paper.

4) Add 100 μL enzyme conjugate to each well,seal the microplate with cover membrance, continue as described in 3.

5) Add 50 μL Substrate A and 50 μL substrate B to each well, shake evenly and incubate at 37 ℃ at dark for 20 min, add 50 μL stop solution to each well.

6) Set microplate reader at 450 nm, adjust blank well as “0”, determine each well “A” value.Also determine at dual-wavelength 450/630 nm, don’t adjust “0”.