Mouse, IgGl

For double immunolabeling with primary antibodies from the same host species, it is not necessary to resort to labeling with monovalent Fab fragments when primary antibodies from the same host species are different isotypes (subclasses) of IgG, for instance such as mouse IgGl and mouse IgG3. In these cases, isotype-specific antibodies may be used to distinguish between the two primary antibodies... 

Concentrate the mouse IgGl or IgG2a antibody to approx 10-15 mg/mL using the Amicon concentration cell and dialyze into 0.02M TE8 buffer. 

Mariam, M., Camagna, M., Tarditi, L and Seccamam, E. (1991) A new enzymatic method to obtain high yield F(ab)2 suitable for clinical use from mouse IgGl. Mol Immunol 28,69—77... 

Basiliximab also binds to the IL-2 receptor alpha chain on activated lymphocytes but is a chimeric human-mouse IgGl. Its indication is identical to that of daclizumab however, basiliximab is given in two bolus or intravenous infusions, the first administered no more than 2 hours prior to transplant and the second 4 days after the transplant procedure. 

Add the detection antibody, 50 pl/well, diluted in PBS/1% BSA. Use either rat anti-mouse IgGl-HRP 1 1000 dilution or rat anti-mouse IgG2a-HRP 1 1000 dilution. 

Chimeric, 10% mouse IgGl, 2% mouse IgG2a 6% Fusion molecules or conjugates... 

Medesan C, Matesoi D, Radu C, Ghetie V, Ward ES. Delineation of the amino acid residues involved in transcytosis and catabolism of mouse IgGl. J Immunol 1997 158(5) 2211-7. 

Figure 19 shows results of adsorption-elution of mouse IgGl monoclonal antibodies on immobilized Procion Red HE-3B. Purity of antibodies is of about 80-85%, few contaminants being still present in the eluted fraction. This separation recipe works also with IgG2 monoclonal antibodies from mouse, however, sodium chloride concentration must be defined case by case. 

Fig. 5. Precipitation of antigen (mouse IgGl) (well 1) by goat antibody to the same antigen (wells 2,5) is inhibited by the Fab fragment of antibody to the same antigen (well 7), but not by Fab from another antibody (to mouse lymphoma EL4) (well 3). Fc from antibody to mouse IgGl (well 6) and rabbit IgG (well 4) likewise fail to inhibit the precipitation.

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