Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (AB170192)

Flow cytometry overlay histogram showing HepG2 cells stained with ab10433 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab10433) (1x10⁶ in 100μl at 5.0μg/ml) for 30 min at 22°C.

The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (ab150117) preadsorbed was incubated at 1/4000 for 30 min at 22°C.

Isotype control antibody (black line) was Mouse IgG2b, kappa monoclonal [7E10G10] (ab170192) used under the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• Flow Cyt

Unknown

Flow Cytometry - Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (AB170192)

Flow cytometry experiments with 4% PFA fixed Hela (human adenocarcinoma), 653s (mouse myeloma), H4IIE (rat hepatoma) and H9C2 (rat myoblast)were performed with the IgG2b Isotype Control Antibody (red) and no primary antibody negative control (black). An Alexa Fluor® 488 conjugate with isotype specificity to the mouse antibody was used as a secondary antibody.

The isotype control at 1 ug/mL shows no higher signal than the no primary negative control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (AB170192)

Immunofluorscent analysis of NIH 3T3, COS7, H9C2 and HeLa cells using ab170192 at 1μg/ml (top); no primary antibody negative control (middle), and ab14705 as a positive control (bottom). An Alexa Fluor® 488 conjugate with isotype specificity to the mouse antibody was used as a secondary antibody. The isotype control at 1 ug/mL shows no higher signal than the no primary negative control.

• ELISA

Supplier Data

ELISA - Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (AB170192)

An isotyping ELISA was performed by coating a 96-well plate with 1.25 ug/mLab170192 and detecting with Alexa Fluor® conjugates specific to mouse IgG1, IgG2a, IgG2b, IgG3, IgM and heavy and light chains (H&L) of IgG. This experiment verifies that the primary antibody's isotype is correct and that it is successfully bound by the secondary antibody.