The Biomat 96 Well Plates for Mouse Monoclonal Antibodies Isotyping product is a 96 well microplate coated with:
- goat anti mouse heavy chain (IgG subclasses 1, 2a, 2b, 3 and IgA and IgM classes)
- goat anti mouse light chain (kappa and lambda)
for qualitative isotype determination of mouse immunoglobulins. This plate enables accurate identification of mouse immunoglobulin isotypes from purified antibodies by capture Enzyme Linked Immunosorbent Assay (ELISA).
Isotyping involves determining the class (e.g. IgG vs IgA) and subclass (e.g. IgG2a vs IgG2b) of a monoclonal antibody: this is a critical step in antibody production as it is necessary for choosing an appropriate purification and modification method for the antibody.
The microplate can be used for performing an ELISA strip-well plate with individual wells precoated with goat anti-mouse heavy chain capture antibody (anti IgG1, IgG2a, IgG2b, IgG3, IgA, IgM) and goat anti-mouse light chain capture antibody (kappa and lambda). A mouse monoclonal antibody sample applied to the wells can be isotyped within an hour and half. Results can be evaluated qualitatively by visual inspection or quantitatively by measuring the absorbance at 450 nm (i.e. if using a goat anti-mouse HRP conjugate as detector).
Features of Biomat coated plates for mouse monoclonal antibody isotyping :
- eight well strip format allows a convenient partial use of the plate; use one strip (column) for each sample (12 samples per plate)
- characterize specific antibodies for six different subclasses and two different light-chain types (see figure below)
- accurate reactivity and specificity characterization with samples containing at least 0.05-1 µg/ml of IgG and 0.1-0.2 µg/ml of IgA and IgM
- high compatibility using samples coming from hybridoma cell culture supernatant, ascites fluid, or purified antibodies
- no special equipment is needed to process the microplate; assess results qualitatively evaluated visually or measured quantitatively using an ordinary ELISA plate reader
Materials provided upon request
Biomat can provide the following reagents to carry out the ELISA isotyping assay:
Individually pouched 96-well microplates with 12 removable 8-well strips:
8-well strip coating
Affinity purified goat anti mouse Igs Fc specific (IgG subclasses 1, 2a, 2b, 3 and IgA and IgM classes) and affinity purified goat anti mouse light chain (kappa and lambda) are coated using 100 µl/well, vertically from position A to position H. The strips are post-coated (blocked) for low non specific binding and long-term stability.
Specificity and reactivity
Mouse monoclonal antibody isotyping coated surfacesee Specificity and reactivity technical note
All wells show a CV% less than 10 when used as a sandwich of mouse Igs in an ELISA format using goat anti mouse Igs (G/A/M)-HRP as detector and TMB as substrate.
Storage and Stability
The microplates, if unopened, are stable refrigerated until the expiration date printed on the label. If opened, store in closed pouch with desiccant and use within the expiration date.