Alexa Fluor® 488 Anti-Acetyl Coenzyme A Carboxylase antibody [EPR23235-147]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Acetyl Coenzyme A Carboxylase antibody [EPR23235-147] (AB326244)

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and ACACA knockout HAP1 stained with ab326244 (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 (ab326244) (1x 10⁶ in 100μl at 0.04 μg/ml (1/12500)) for 30min at 22°C.

Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HAP1 WT cells (black line) and HAP1-ACACA KO cells (grey line) was also included.

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

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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Acetyl Coenzyme A Carboxylase antibody [EPR23235-147] (AB326244)

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 ab326244 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). PBMCs were incubated for 30 min at 4°C 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 (ab326244) or Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶ 100μl at 0.2 μg/ml (1/2500)) for 30min on ice. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

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