Mouse Monoclonal Antibodies Isotyping ELISA Coated Surface 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.

With Biomat 96-Well ELISA Plate you can be isotyped, within an hour and half, one or more Mouse Monoclonal Antibody samples.

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). 

Results can be evaluated both qualitatively by visual inspection and quantitatively, by measuring the absorbance at 450 nm (i.e. if using a goat anti-mouse HRP conjugate as detector).

Biomat’s Coated 96-Well Plates for Isotyping ELISA Assay

This is the available configuration of Mouse Mab Isotyping  96-Well Coated Plate products by Biomat.

Click on the configuration link to scroll down the page to the relative section.

(on request Biomat can supply microplates for fluorescent and chemiluminescent assays)

Coated 96-Well Plates for Mouse Mab Isotyping ELISA Test main features

Biomat’s Coated 96-Well Plates for Mouse Monoclonal Antibodies Isotyping ELISA Assay  have the following characteristics:

  • Coated with goat anti mouse heavy chain (IgG subclasses 1, 2a, 2b, 3 and IgA and IgM classes) and goat anti mouse light chain (kappa and lambda)
  • Ready to use
  • Manufactured under ISO:9001 guidelines
  • Post-coated (blocked) for low non specific binding and long-term stability
  • 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
  • All lots are tested for uniformity and reproducibility
  • Certificate of Quality is released for every lot
  • For Research Use Only

Biomat can provide the following reagents to carry out the ELISA Isotyping Assay:

Mouse Mab Isotyping ELISA Product specifications

Coating – 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.

Specificity and Reactivity

Six mouse monoclonal antibodies were analyzed to prove the specificity along with the reactivity of the strips.

see Specificity and reactivity technical note

Uniformity

All wells of the 96-Well Coated for Mouse Monoclonal Antibodies Isotyping ELISA product shows 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 at 2-8°C until the expiration date printed on the label. If opened, store in closed pouch with desiccant and use within the expiration date.