IgM ELISA 试剂盒

Size / Description
• Pathogen antigen coated 96-well Microplate
• 96 wells (12 strips x 8 wells) coated with pathogen antigen
• Wash Buffer Concentrate (20X)
• 25 ml of 20X concentrated solution
• Positive Control
• 1 vial of Positive Control sample containing anti-pathogen antigen antibody
• Biotinylated Anti-Human Ig(A/G/M)
• 1 vial of solution
• Biotinylated Anti-Positive Control
• 1 vial of solution
• HRP-Streptavidin concentrate
• 1 vial of solution
• TMB One-Step Substrate Reagent
• 12 ml of 3,3,5,5'-tetramethylbenzidine (TMB buffer solution)
• Stop Solution
• 8 ml of 0.2 M sulfuric acid
• Assay Diluent B
• 15 ml of 5X concentrated buffer
• 5X Sample Diluent
• 25 ml of 5X diluent buffer, 0.5% proclin 300 as preservative

• Microplate reader capable of measuring absorbance at 450 nm
• Shaker
• Pipettes capable of accurately delivering 2 µL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Graduated cylinders: 100 mL and 1 liter
• Distilled or deionized water
• Absorbent paper

1. Prepare all reagents, samples and standards as instructed.
2. Add 100 μL positive control, or sample to each well. Incubate 1 hour at room temperature.
3. Add 100 μL prepared Biotinylated Anti-Positive Control and Biotinylated Anti-Human Ig(A/G/M) Antibody into respective wells. Incubate 1 hour at room temperature.
4. Add 100 μL prepared HRP-Streptavidin solution to each well. Incubate 30 minutes at room temperature.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Gene Symbols: IgM

Background: This in-vitro ELISA kit employs an indirect ELISA method. Standard 96-well plates are coated with the pathogen antigen, which combines with the corresponding antibody present in a sample and positive control, which is used as a calibration curve for interpretation purposes. When a biotinylated secondary anti-human IgA/G/M antibody is added, a complex forms between the biotinylated secondary antibody and human antibody bound to the protein-coated plate. A TMB substrate is added, and a blue color is generated. The depth of color is relative to the amount of the human anti-pathogen antigen antibody present. The stop solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

The Positive Control consists of an anti-pathogen antigen antibody. A biotinylated anti-positive control antibody is added to the positive control wells and treated identically to the samples. The intensity of the resulting color (measured at 450 nm) is used as a calibration curve for interpretation purposes in the different assays.< h4>Kit Components< table class="hasborder">ComponentSize / DescriptionPathogen antigen coated 96-well Microplate96 wells (12 strips x 8 wells) coated with pathogen antigenWash Buffer Concentrate (20X)25 mL of 20X concentrated solutionPositive Control1 vial of Positive Control sample containing anti-pathogen antigen antibodyBiotinylated Anti-Human Ig(A/G/M)1 vial of solutionBiotinylated Anti-Positive Control1 vial of solutionHRP-Streptavidin concentrate1 vial of solutionTMB One-Step Substrate Reagent12 mL of 3,3,5,5"-tetramethylbenzidine (TMB buffer solution)Stop Solution8 mL of 0.2 M sulfuric acidAssay Diluent B15 mL of 5X concentrated buffer5X Sample Diluent25 mL of 5X diluent buffer, 0.5 % proclin 300 as preservative< h4>Additional Materials Required< div class="desc-content">

• Microplate reader capable of measuring absorbance at 450 nm
• Shaker
• Pipettes capable of accurately delivering 2 µL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Graduated cylinders: 100 mL and 1 liter
• Distilled or deionized water
• Absorbent paper

Protein Name (Synonyms): IgM, Human Coronavirus OC43 Hemagglutinin Esterase (HE)