Peroxidase affinipure bovine anti goat igg h l

Example 3

Standard ELISA procedures were performed for the determination of antigen binding affinity using the following sets of reagents. ELISA plates were coated with BSA-SSEA4 at a 1:1000 dilution. Samples of serially-diluted hybridoma supernatants or transient transfection samples were added to each well, followed by thorough washing. The following secondary antibodies were used to detect muMAbs and huMAbs bound to the antigen on the plate: For muMAb1, AffiniPure Goat Anti-Mouse IgG, Fcγ Subclass 1 Specific (Jackson ImmunoResearch #115-005-205) at a 1:2,500 dilution, for muMAb2, AffiniPure Goat Anti-Mouse IgG, Fcγ Subclass 3 Specific (Jackson ImmunoResearch #115-005-209) at a 1:2,500 dilution, for all humanized antibodies, AffiniPure Goat Anti-Human IgG (H+L) (Jackson ImmunoResearch #109-005-088) at a 1:2,500 dilution.

Plate-bound secondary antibodies were detected using Peroxidase AffiniPure Bovine Anti-Goat IgG (H+L) (Jackson ImmunoResearch #805-035-180) at a 1:6,000 dilution and developed as set forth above.

Example 3

Standard ELISA procedures were performed for the determination of antigen binding affinity using the following sets of reagents. ELISA plates were coated with BSA-SSEA4 at a 1:1000 dilution. Samples of serially-diluted hybridoma supernatants or transient transfection samples were added to each well, followed by thorough washing. The following secondary antibodies were used to detect muMAbs and huMAbs bound to the antigen on the plate: For muMAb1, AffiniPure Goat Anti-Mouse IgG, Fcγ Subclass 1 Specific (Jackson ImmunoResearch #115-005-205) at a 1:2,500 dilution, for muMAb2, AffiniPure Goat Anti-Mouse IgG, Fcγ Subclass 3 Specific (Jackson ImmunoResearch #115-005-209) at a 1:2,500 dilution, for all humanized antibodies, AffiniPure Goat Anti-Human IgG (H+L) (Jackson ImmunoResearch #109-005-088) at a 1:2,500 dilution.

Plate-bound secondary antibodies were detected using Peroxidase AffiniPure Bovine Anti-Goat IgG (H+L) (Jackson ImmunoResearch #805-035-180) at a 1:6,000 dilution and developed as set forth above.