Product name HAV-IgM Elisa kit for Research Use Only
This kit uses capture ELISA method to detect anti-HAV IgM. The
purified mouse anti-human IgM (μchain) antibody is coated on the
solid phase of multi-wells. A conjugate HAV-Ag is added to the
coated wells after diluted sample added and incubated. Then
horseradish peroxidase labeled HAV-Ag is added. If HAV-IgM is
present in the sample, a complex of Anti-μ-chain-HAV-IgM –HAV-Ag
-labeled with HRP will form. Wash wells to remove other unbounded
serum components, incubate with substrate (TMB) to form a colored
product, and measure the absorbance at 450nm to indicate the
presence or absence of HAV-IgM in the sample. The test is special,
sensitive, reproducible and easy to operate.
Main Component
1. Anti-μ-chain Coated Microwell Plate
2. Enzyme Conjugant (HAVAg-HRP)
3. Conjugate HAV-Ag
4. Negative Control Serum
5. Positive Control Serum
6. 20 X Wash Buffer (dilution prior to use)
7. Substrate A
8. Substrate B
9. Stop Solution
10. Plastic Bag
11. Seal Paper
12. Manual
TEST PROCEDURE
1. Bring ELISA Kit for Antibody IgM to Hepatitis A Virus (all
reagents), and samples to room temperature before use
(approximately 30 minutes).
2. Dilute concentrated wash buffer 1:19 with ddH2O.
3. Dilute the sample (1:1000) with physiological saline.
4. For each test, set one blank, two positive and three negative
controls. Add 100μl positive and negative control serum into
positive and negative control wells respectively.
5. Add 100μl diluted sample into other test wells.
6. Cover wells with seal paper, then incubate 30 minutes at 37°C.
7. Discard the liquid in all wells and fill the wells with wash
solution. Lay aside for 15 seconds, discard the liquid in all wells
and fill the wells with wash solution. Repeat 5 times and dry wells
after last wash.
8. Add 50 μl conjugate HAV-Ag in each well except the blank.
9. Addμl 50 μl Enzyme Conjugant in each well except the blank
10.Cover wells with seal paper, then incubate 30 minutes at 37°C.
11. Repeat step 7.
12.Add 50 μl substrate A and B respectively to each well,mix gently
protected from light and incubate 15 minutes at 37°C.
13. Add 50μl of stop solution into each well to stop the reaction,
including blank well.
14. Measure the absorbance at 450 nm against the blank, or measure
the absorbance at 450 nm/630-690 nm.
