Anti-SATB2 antibody [EPNCIR130B] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SATB2 antibody [EPNCIR130B] - BSA and Azide free (AB212176)

This data was developed using ab133328, the same antibody clone in a different buffer formulation.Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon cancer tissue sections labeling SATB2 with Purified ab133328 at 1 : 100 dilution (1.57 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control : PBS instead of the primary antibody.Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SATB2 antibody [EPNCIR130B] - BSA and Azide free (AB212176)

This data was developed using ab133328, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labelling SATB2 with unpurified ab133328 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-SATB2 antibody [EPNCIR130B] - BSA and Azide free (AB212176)

This data was developed using ab133328, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-SATB2 antibody [EPNCIR130B] (<a href='/en-us/products/primary-antibodies/satb2-antibody-epncir130b-ab133328'>ab133328</a>) at 1/1000 dilution

Lane 1:

HT-1080 cell lysate at 10 µg

Lane 2:

Saos-2 cell lysate at 10 µg

Lane 3:

Human fetal brain lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 83 kDa

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• WB

Lab

Western blot - Anti-SATB2 antibody [EPNCIR130B] - BSA and Azide free (AB212176)

This data was developed using ab133328, the same antibody clone in a different buffer formulation.

Lanes 1 - 2 : Merged signal (red and green). Green - ab133328 observed at 83 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab133328 was shown to specifically react with SATB2 in wild-type HAP1 cells as signal was lost in SATB2 knockout cells. Wild-type and SATB2 knockout samples were subjected to SDS-PAGE. ab133328 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-SATB2 antibody [EPNCIR130B] (<a href='/en-us/products/primary-antibodies/satb2-antibody-epncir130b-ab133328'>ab133328</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

SATB2 knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 83 kDa

Observed band size: 83 kDa

false

• WB

Lab

Western blot - Anti-SATB2 antibody [EPNCIR130B] - BSA and Azide free (AB212176)

This data was developed using ab133328, the same antibody clone in a different buffer formulation.

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

Freshly made lysate is highly recommended to minimize protein degradation.

All lanes:

Western blot - Anti-SATB2 antibody [EPNCIR130B] (<a href='/en-us/products/primary-antibodies/satb2-antibody-epncir130b-ab133328'>ab133328</a>) at 1/10000 dilution

Lane 1:

Saos-2 (Human osteosarcoma epithelial) whole cell fresh lysate at 10 µg

Lane 2:

Saos-2 (Human osteosarcoma epithelial) whole cell lysate at 10 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: 83 kDa

Observed band size: 83 kDa

false

Exposure time: 20s