Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control

• ELISA

Supplier Data

ELISA - Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (AB170190)

Fluorescence ELISA.

An isotyping ELISA was performed by coating a 96-well plate with 1.25 ug/mL of the IgG1 Isotype Control Antibody 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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (AB170190)

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and VIM knockout HAP1 cells stained with ab8069 (magenta line). The cells were fixed with 80% methanol (5 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 (ab8069) (1x10⁶ in 100 μl at 0.04 μg/ml) for 30 min at 22°C.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/4000 for 30 min at 22°C.

Isotype control antibody was mouse IgG1κ (ab170190) used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line, VIM knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (AB170190)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (AB170190)

Immunocytochemistry (ICC) experiments with NIH-3T3 (mouse sarcoma), COS7 (monkey kidney fibroblast), H9C2 (rat myoblast) and Hela (human adenocarcinoma) were performed with the IgG1 Isotype Control Antibody (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.

• Flow Cyt

Unknown

Flow Cytometry - Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (AB170190)

Flow cytometry experiments with 4% PFA fixed HeLa (Human adenocarcinoma), 653s (mouse myeloma), H4IIE (rat hepatoma) and H9C2 (rat myoblast) were performed with the IgG1 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.