Bio-Rad Mouse Typer Isotyping Kit - Manual (Page 3)

Section 1
Introduction
1.1 Background
The growth and widespread use of mouse monoclonal
antibody technology have created a need for a fast, accurate, and
simple means of determining immunoglobulin class and sub-class.
Several classes of mouse monoclonal antibody, all structurally
different depending on their heavy chain composition, have been
described.
1 Identification is essential since chemical and biological
properties of the various classes are unique. They differ in their
solubility and electrophoretic properties, in their susceptibility to
cleavage enzymes, and in their reactivity with protein A.
2,3
The Mouse Typer sub-isotyping kit is an extremely
convenient kit for identifying mouse immunoglobulin class and
sub-class in tissue culture supernatant and ascitic fluids. The
ELISA based kit uses a panel of ultra pure reagents to determine
mouse sub-isotypes: IgG
1
, IgG2
a
, IgG2
b
, IgG
3
, IgM, IgA,
χ
chain,
and
λ
chain. It comes complete with all the essential reagents for
800 extremely sensitive tests (100 typings). The included protocol
and troubleshooting guide guarantee efficient and accurate assays.
Determining a mouse monoclonal antibody’s class with the
Mouse Typer kit is fast and simple. Antigen is first adsorbed to a
microplate, then treated with specific monoclonal antibody. Bound
monoclonals are separately reacted with each of the Mouse Typer
rabbit anti-mouse panel reagents. Immunoglobulin class and sub-class
are then determined with Bio-Rad’s goat anti-rabbit (H + L)
horseradish peroxidase (HRP) conjugate and peroxidase substrate
system. Color development occurs immediately, with positive wells
having absorbances 3-6 times that of corresponding negatives. Using
Bio-Rad’s Model 3550 or Model 3550-UV Microplate Reader, results
are easily quantitated in less than 60 seconds.
1
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