Recombinant Anti-STAT6 antibody [E265] - BSA and Azide free (ab247230)

This data was developed using ab32108, the same antibody clone in a different buffer formulation.

Confocal image showing cytoplasmic staining in Raji cells. Immunocytochemistry/immunofluorescence analysis of Raji (human Burkitt's lymphoma B lymphocyte) cells labelling STAT6 with ab32108 at 10 µg/mL. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as a secondary antibody at 2 µg/mL. Cells were counterstained with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).

This data was developed using ab32108, the same antibody clone in a different buffer formulation.

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)

Lane 2: STAT6 knockout HAP1 whole cell lysate (20 µg)

Lane 3: HeLa whole cell lysate (20 µg)

Lane 4: Raji whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab32108 observed at 94 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab32108 was shown to specifically react with STAT6 in wild-type HAP1 cells as signal was lost in STAT6 knockout cells. Wild-type and STAT6 knockout samples were subjected to SDS-PAGE. ab32108 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

This data was developed using ab32108, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling STAT6 with purified ab32108 at 1/100 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).

This data was developed using ab32108, the same antibody clone in a different buffer formulation.
Purified ab32108 at 1/50 dilution (2µg) immunoprecipitating STAT6 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab32108 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32108 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 100 kDa

This data was developed using ab32108, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: Lane 1. 7 seconds ; Lane 2. 3 seconds

This antibody recognizes three isoforms of STAT6, isoform 1 (110kDa), isoform 3 (81kDa), which is consistent with the Uniprot annotation.