Recombinant Anti-SATB1 antibody [EPR3951] - BSA and Azide free (ab239944)

This data was developed using ab239944, the same antibody clone in a different buffer formulation.
Purified ab239944 at 1:20 dilution (0.6µg) immunoprecipitating SATB1 in Rat thymus lysate.
Lane 1 (input): Rat thymus lysate 10 µg
Lane 2 (+): ab239944 + Rat thymus lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109122 in Rat thymus lysate.
VeriBlot for IP Detection Reagent (HRP)(ab131366) (1:5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: kDa

This data was developed using ab239944, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling SATB1 with Purified ab239944 at 1:20 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). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using ab239944, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling SATB1 with Purified ab239944 at 1:50 dilution (2.5µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using ab109122, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling SATB1 with Purified ab109122 at 1:500 dilution (0.246 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

This data was developed using ab109122, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling SATB1 with Purified ab109122 at 1:500 dilution (0.246 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

This data was developed using ab109122, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling SATB1 with Purified ab109122 at 1:500 dilution (0.246 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

This data was developed using the same antibody clone in a different buffer formulation (ab109122).

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: SATB1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: Jurkat whole cell lysate (20 µg)
Lane 4: THP1 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab109122 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab109122 was shown to specifically react with SATB1 in wild-type HAP1 cells as signal was lost in SATB1 knockout cells. Wild-type and SATB1 knockout samples were subjected to SDS-PAGE. Ab109122 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.