Biovantion Inc.
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Human High Quality HEV IgG Elisa Kit 96Test/Kit
Diagnositc Kit for IgG Antibody to Hepatitis E Virus(ELISA)
Catalog No.: BE401A
1. PRINCIPLE
This kit employs solid phase, indirect ELISA method for detection of IgG antibodies to HEV (anti-HEV) in serum or plasma with two-step incubation procedure. Polystyrene microwell are pre-coated with purified activated recombinant HEV antigen. The HRP conjugated mouse anti human IgG (r chain) monoclonal antibody serves as tracer. TMB is substrate for HRP. The enzyme reaction with substrate TMB produces a color change, and the intensity of the absorbance at 450 nm indicates the presence or absence of Anti-HEV antibodies IgM in the sample. The test is specific, sensitive, reproducible and easy to operate. It is for blood screen of HEV infection.
2. MATERIALS PROVIDED
1. Antigen Coated Microwell Plate 1 block (96wells)
2. Specimen Diluent 1 bottle (12ml)
3. Negative Control 1 vial (1ml)
4. Positive Control 1 vial (1ml)
5.20 X Wash Buffer (dilution prior to use) 1 bottle (30ml)
6.Enzyme Conjugant (anti human IgG -HRP) 1 bottle (12ml)
7. Substrate A 1 bottle (6ml)
8. Substrate B 1 bottle (6ml)
9. Stop Solution(2M H2SO4) 1 bottle (6ml)
10. Plastic Bag 1 bag
11. Seal Paper 3 pieces
12. Manual 1 each
TEST PROCEDURE
1. Bring ELISA Kit for Antibody IgG to Hepatitis E Virus (all reagents), and Specimens to room temperature before use (approximately 30 minutes).
2. Dilute concentrated wash buffer 1:19 with ddH2O.
3. 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.
4. Add 100 μl sample diluent in each other test wells, then at 10 μl test serum into test wells. Pipet up and down to mix the samples well.
5. Cover wells with seal paper, then incubate 30 minutes at 37°C.
6. 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.
7. Add 100 μl Enzyme Conjugant in each well except the blank.
8. Cover wells with seal paper, then incubate 30 minutes at 37°C.
9. Repeat step 6.
10. Add 50μl substrate A and B respectively to each well including the blank well. Mix gently, protected from light and incubate 15 minutes at 37°C.
11. Add 50μl of stop solution into each well to stop the reaction, including blank well.
12. Measure the absorbance at 450 nm against the blank, or measure the absorbance at 450 nm/630-690 nm.
