Mouse Ig Isotyping ELISA Kit

Detection Principle:

This kit uses double antibody sandwich enzyme-linked immunosorbent assay technology for qualitative identification. Monoclonal antibodies specific for mouse immunoglobulin subtypes were pre coated on high affinity enzyme labeled plates. The positive control sample, negative control sample, sample to be tested and horseradish peroxidase conjugated detection antibody were added into the wells of enzyme plate. After incubation, the immunoglobulin in the sample was combined with the solid-phase antibody and detection antibody. After thorough washing to remove unbound substances, TMB chromogenic substrate was added and incubated in the dark to develop color. The reaction was stopped by adding stop solution, and the absorbance value was measured at 450 nm wavelength (correction wavelength 570 - 630 nm).

Detection Type: double antibody sandwich method

Form: pre coated 96 well plate

Test Sample Type: cell supernatant, serum, plasma

Loading Amount: 100 μ L

Kit Components: A copy of pre coated 96 well plate, positive control, Ig isotyping detection antibody, detection buffer, TMB chromogenic substrate, washing solution, termination solution, plate sealing membrane and instructions.

Storage Method: 2-8 ℃

Plate Layout:

abbreviations:

pc- positive control positive control   nc- negative control negative control   spl.1-10- sample 1-10 #1-10 samples

Background:

Antibody typing pair hybridization tumor cell preparation is very critical and useful. This kit can identify 6 immunoglobulin heavy chain subtypes and 2 light chain subtypes in mice, which are IgA, IgM, IgG1, IgG2a, IgG2b, IgG3, & kappa; Chain and λ Chain. It can accurately and specifically identify the types of heavy and light chains produced by hybridoma cells, and identify whether they are monoclonal antibodies. Therefore, this kit is a very useful tool to distinguish and identify each potential clone. Because the chemical and biological characteristics of various subtypes are different, it is essential to identify them. They differ in solubility, electrophoretic properties, sensitivity to lyases, and reactivity with protein A. Therefore, identifying the classes and subclasses of monoclonal antibodies is very useful for selecting the best way of immunoglobulin purification. For example, purification of mouse IgA and IgM by molecular weight (such as gel chromatography) or immunoaffinity separation column is the best method. Mouse IgG2a and IgG2b can be purified by solid-phase protein A at ph7-8, while mouse IgG1 can be purified by solid-phase protein A at ph8-9Κ Chain immunoglobulins can be purified by solid-phase protein L.