Mouse IgG2a ELISA Kit

Catalog No.: BEK1296

Size: 96T

Range: 390 pg/ml - 25,000 pg/ml

Sensitivity < 190 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of IgG2a in mouse serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Immunoglobulin G (IgG) is the most abundant Ig isotype in serum, making up approximately 80% of all serum Immunoglobulins. In mice, the IgG subclasses are defined IgG1, IgG2a/c, IgG2b, and IgG3.  The IgG2a allele is expressed in e.g., Balb/c mice. IgG2a is known as the most active IgG subclass in mice due to its high A/I ratio.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- IgG2a polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- IgG2a polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the IgG2a amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IgG2a can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-mouse IgG2a antibody
2. Lyophilized mouse IgG2a standards: 2 tubes (50 ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-mouse IgG2a antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L