Anti-SATB1 antibody [EPR3951] - BSA and Azide free

• Flow Cyt

Lab

Flow Cytometry - Anti-SATB1 antibody [EPR3951] - BSA and Azide free (AB239944)

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).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SATB1 antibody [EPR3951] - BSA and Azide free (AB239944)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SATB1 antibody [EPR3951] - BSA and Azide free (AB239944)

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. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SATB1 antibody [EPR3951] - BSA and Azide free (AB239944)

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. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SATB1 antibody [EPR3951] - BSA and Azide free (AB239944)

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. PBS instead of the primary antibody was used as the negative control.

• IP

Lab

Immunoprecipitation - 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

All lanes:

Immunoprecipitation - Anti-SATB1 antibody [EPR3951] (<a href='/en-us/products/primary-antibodies/satb1-antibody-epr3951-ab109122'>ab109122</a>)

Predicted band size: 85 kDa

false

• WB

Supplier Data

Western blot - Anti-SATB1 antibody [EPR3951] - BSA and Azide free (AB239944)

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.

All lanes:

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

Predicted band size: 85 kDa

false

• WB

Unknown

Western blot - Anti-SATB1 antibody [EPR3951] - BSA and Azide free (AB239944)

All lanes:

Western blot - Anti-SATB1 antibody [EPR3951] (<a href='/en-us/products/primary-antibodies/satb1-antibody-epr3951-ab109122'>ab109122</a>) at 1/1000 dilution

Lane 1:

LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

THP-1 (Human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 3:

Mouse liver lysate at 20 µg

Lane 4:

Mouse thymus lysate at 20 µg

Lane 5:

Rat thymus lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 85 kDa

false

• WB

Unknown

Western blot - Anti-SATB1 antibody [EPR3951] - BSA and Azide free (AB239944)

All lanes:

Western blot - Anti-SATB1 antibody [EPR3951] (<a href='/en-us/products/primary-antibodies/satb1-antibody-epr3951-ab109122'>ab109122</a>) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia T lymphocyte) nuclear extract lysate at 20 µg

Lane 2:

Jurkat (Human T cell leukemia T lymphocyte) without nuclear extract lysate at 20 µg

Lane 3:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 85 kDa

false