Anti-FOXO3A antibody [EPR29854-587] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXO3A antibody [EPR29854-587] - BSA and Azide free (AB325946)

This data was developed using ab323467, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling FOXO3A with ab323467 at 1/500 (1.004 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon (PMID : 23828518).
The section was incubated with ab323467 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-FOXO3A antibody [EPR29854-587] - BSA and Azide free (AB325946)

This data was developed using ab323467, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human adenocarcinoma of stomach tissue labeling FOXO3A with ab323467 at 1/500 (1.004 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human adenocarcinoma of stomach (PMID : 33795838).
The section was incubated with ab323467 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-FOXO3A antibody [EPR29854-587] - BSA and Azide free (AB325946)

This data was developed using ab323467, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling FOXO3A with ab323467 at 1/500 (1.004 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung cancer.
The section was incubated with ab323467 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-FOXO3A antibody [EPR29854-587] - BSA and Azide free (AB325946)

This data was developed using ab323467, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling FOXO3A with ab323467 at 1/500 (1.004 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.
The section was incubated with ab323467 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-FOXO3A antibody [EPR29854-587] - BSA and Azide free (AB325946)

This data was developed using ab323467, the same antibody clone in a different buffer formulation.

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

In lanes 1-3 the lysates were stored at -80°C prior to Western Blotting. The bands beneath the target band (82 kDa) are likely to be degradation products. In lane 4 To minimize protein degradation cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the higher MW band at approximately 150 kDa (in lanes 1-3) is unknown.

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

Exposure time : Lanes 1-3 : 180 seconds lane 4 : 37 seconds

All lanes:

Western blot - Anti-FOXO3A antibody [EPR29854-587] (<a href='/en-us/products/primary-antibodies/foxo3a-antibody-epr29854-587-ab323467'>ab323467</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting FOXO3A whole cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervical adenocarcinoma epithelial cell) 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 1/100000 dilution

Observed band size: 82 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-FOXO3A antibody [EPR29854-587] - BSA and Azide free (AB325946)

This data was developed using ab323467, the same antibody clone in a different buffer formulation.

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

To minimize protein degradation cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

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

Exposure time : Lanes 1 3 : 10 seconds lanes 2 4-6 : 37 seconds

All lanes:

Western blot - Anti-FOXO3A antibody [EPR29854-587] (<a href='/en-us/products/primary-antibodies/foxo3a-antibody-epr29854-587-ab323467'>ab323467</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 6:

SH-SY5Y (human neuroblastoma epithelial cell) 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 1/100000 dilution

Observed band size: 82 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-FOXO3A antibody [EPR29854-587] - BSA and Azide free (AB325946)

This data was developed using ab323467, the same antibody clone in a different buffer formulation.

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

The identity of the lower MW band at approximately 15 kDa (in lane 6) is unknown.

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

Exposure time : Lanes 1-2 : 37 seconds lanes 3-6 : 180 seconds

All lanes:

Western blot - Anti-FOXO3A antibody [EPR29854-587] (<a href='/en-us/products/primary-antibodies/foxo3a-antibody-epr29854-587-ab323467'>ab323467</a>) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse pancreas tissue lysate at 20 µg

Lane 4:

Mouse heart tissue lysate at 20 µg

Lane 5:

Mouse brain tissue lysate at 20 µg

Lane 6:

Rat brain tissue 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 1/100000 dilution

Observed band size: 82 kDa,36 kDa

false