Anti-FOXA1 antibody [RM1119] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell, Left ) / PC-3 (human prostate adenocarcinoma epithelial cell, Right ) cells labelling FOXA1 with ab317046 at 1/5000 dilution (0.01 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : HCT 116

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) PC-3 (human prostate adenocarcinoma epithelial cell) cell pellet (B) HCT-116 (human colorectal carcinoma epithelial cell) cell pellet tissue labeling FOXA1 with ab317046 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) PC-3 cell pellet, no staining on (B) HCT-116 cell pellet.
The section was incubated with ab317046 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling FOXA1 with ab317046 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human breast cancer.
The section was incubated with ab317046 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling FOXA1 with ab317046 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human prostatic hyperplasia.
The section was incubated with ab317046 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-3 (human prostate adenocarcinoma epithelial cell) cells labelling FOXA1 with ab317046 at 1/2000 (0.261 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear staining in PC-3 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : HCT 116.
The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling FOXA1 with ab317046 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on rat colon.
The section was incubated with ab317046 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling FOXA1 with ab317046 at 1/2000 (0.522 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on mouse colon.
The section was incubated with ab317046 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

The lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-FOXA1 antibody [RM1119] (<a href='/en-us/products/primary-antibodies/foxa1-antibody-rm1119-ab317046'>ab317046</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

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 1/100000 dilution

Observed band size: 52 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Negative controls : HCT 116 and SH-SY5Y.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane1, 3-5 : 180 seconds Lane2 : 37 seconds

All lanes:

Western blot - Anti-FOXA1 antibody [RM1119] (<a href='/en-us/products/primary-antibodies/foxa1-antibody-rm1119-ab317046'>ab317046</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 3:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 4:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 5:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

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 1/100000 dilution

Observed band size: 35-52 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

FOXA1 can undergo degradation easily. The lysates were recommended freshly made and used for Western Blotting immediately to minimize protein degradation

Exposure time : Lane 1 : 15 seconds Lane 2 : 37 seconds

All lanes:

Western blot - Anti-FOXA1 antibody [RM1119] (<a href='/en-us/products/primary-antibodies/foxa1-antibody-rm1119-ab317046'>ab317046</a>) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) whole fresh cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

PC-3 (human prostate adenocarcinoma epithelial cell) whole frozen cell lysate at 20 µg with 5% NFDM/TBST

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 49 kDa,52 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [RM1119] - BSA and Azide free (AB317047)

This data was developed using ab317046, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling FOXA1 with ab317046 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human cerebrum.
The section was incubated with ab317046 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins