PE Anti-IDH2 antibody [EPR7577]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - PE Anti-IDH2 antibody [EPR7577] (AB212122)

Flow cytometry scatter plots showing untreated human peripheral mononuclear cells (PBMCs) (top) and PBMCs activated with 5 μg/ml PHA for 18h (bottom), either stained with ab212122 (right) or isotype control (left). The cells were fixed using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab212122) (1x 10⁶ in 100μl at 0.04 μg/ml (1/12500)) for 30min on ice. Isotype control antibody was Rabbit IgG (monoclonal) Phycoerythrin Isotype Control (ab209478) - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 50 mW Yellow/Green laser (561nm) and 585/42 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - PE Anti-IDH2 antibody [EPR7577] (AB212122)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat IDH2 knockout cells labeling IDH2 with ab212122 (green) at 1 μg/ml. Cells were counterstained with ab190573 Anti-Tubulin (Alexa Fluor® 647) at 1 : 250 (2 μg/ml) (magenta). The nuclear counterstain was DAPI (blue).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - PE Anti-IDH2 antibody [EPR7577] (AB212122)

Overlay histogram showing MCF7 cells stained with ab212122 (red line). The cells were fixed with 4% formaldehyde and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab212122, 1/1000 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 50mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - PE Anti-IDH2 antibody [EPR7577] (AB212122)

Flow cytometry overlay histogram showing wild-type Jurkat (green line) and IDH2 knockout Jurkat stained with ab212122 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised 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 (ab212122) (1x 10⁶ in 100μl at 0.008 μg/ml (1/62500)) for 30min at 22°C.

Isotype control antibody was Rabbit IgG (monoclonal) Phycoerythrin Isotype Control (ab209478) in Jurkat WT cells (black line) and Jurkat-IDH2 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Yellow/Green laser (561nm) and 585/42 bandpass filter.