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ELISA Operation Manual iElisa Afla Total

ELISA Operation Manual iElisa Afla Total. Clover Technology Group Inc. 1. Preparation of working. Return all reagents to 2 - 8 °C (35 - 46 °F) immediately after use. Bring all reagents to room temperature (20 - 25 °C / 68 - 77 °F) before use. 2.Preparation of Samples.

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ELISA Operation Manual iElisa Afla Total

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  1. ELISA Operation ManualiElisaAfla Total Clover Technology Group Inc

  2. 1. Preparation of working • Return all reagents to 2 - 8 °C (35 - 46 °F) immediately after use. • Bring all reagents to room temperature (20 - 25 °C / 68 - 77 °F) before use.

  3. 2.Preparation of Samples ①Transfer 5 grams of ground sample into an extraction mixing jar.

  4. ②70 percent methanol/water: Using a graduated cylinder, measure 700 ml of methanol and place it into a clean carboy with spigot. Add 300 ml deionized or distilled water to the methanol and shake vigorously until it is completely mixed. Store this solution at room temperature in a tightly closed container until needed. ③Add 20ml of the (70/30) methanol/water extraction solvent.

  5. Cover the extraction jar and blend on high speed for 10 minutes. • filter entire extract by using a paper filter

  6. 3. Preparation of working solution • 80× Enzyme Conjugate: dilute 80x with Diluent buffer A(10μl+790μl). • - 80×Antibody cross-reactivity: dilute 80x with Diluent buffer A(10μl+790μl). • - wash buffer: dilute 10x with distilled water (50ml+450ml).

  7. Dilute 500μl of the filtrate 750μl of dilute buffer A • Insert a sufficient number of wells into the microwell holder for all standards and samples to be tested

  8. 4. Immunoassay Procwdure • Using a new pipette tip for each standard and sample, pipet 50μl of standards and prepared sample to separate wells. • Add 50μl of enzyme conjugate into each well. • Add 50μl of anti-aflatoxin antibody into each well. • Incubate for 20 minutes at room temperature (37°C)

  9. 5.Washing • Dump the contents of the wells. Turn the wells upside down and tap out on a paper towel until the remaining liquid has been removed. • Using a wash bottle, fill each well with washing buffer .Empty the wells again and remove all remaining liquid. Repeat this step 2 times (total of 4 washes).

  10. 6.Add TMB • Add 150μl of TMB to each well. Incubate for 8 minutes at room temperature (37°C). Cover the wells with a paper towel to protect them from light sources.

  11. 7. Add 50μl of stop solution to each well.

  12. 8. Software installation instructions • 1）Zip file decompress • 2）Double-click • 3）bring in serial Number: 123456

  13. 9. Software Usag • 1）click method to establish a new method • 2）the left :bring in STD Concentration and replicates • 3）The right is for sample After setting click “ok”

  14. Back to main page Under the column corresponding bring in STD OD Under the column corresponding bring in sample OD

  15. for example

  16. cilck you can see the Standard curve

  17. click sample This is the result.

  18. Thanks Clover Technology Group Inc

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