Anti-FOXF1 antibody [EPR27368-79]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXF1 antibody [EPR27368-79] (AB308633)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labelling FOXF1 with ab308633 at 1/1000 (0.52 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression tissue : no staining on rat cardiac muscle. The section was incubated with ab308633 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-FOXF1 antibody [EPR27368-79] (AB308633)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labelling FOXF1 with ab308633 at 1/1000 (0.52 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat lung. The section was incubated with ab308633 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-FOXF1 antibody [EPR27368-79] (AB308633)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labelling FOXF1 with ab308633 at 1/1000 (0.52 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse lung (PMID : 11313147). The section was incubated with ab308633 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-FOXF1 antibody [EPR27368-79] (AB308633)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling FOXF1 with ab308633 at 1/1000 (0.52 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression tissue : no staining on mouse cardiac muscle (PMID : 11313147). The section was incubated with ab308633 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

• IP

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Immunoprecipitation - Anti-FOXF1 antibody [EPR27368-79] (AB308633)

FOXF1 was immunoprecipitated from 0.35 mg Mouse lung tissue lysate with ab308633 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308633 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse lung tissue lysate
Lane 2 : ab308633 IP in Mouse lung tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308633 in mouse lung tissue lysate

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

Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-FOXF1 antibody [EPR27368-79] (ab308633) at 1/1000 dilution

All lanes:

Mouse lung tissue lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 40 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-FOXF1 antibody [EPR27368-79] (AB308633)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : A375 (PMID : 29162563). The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29162563, 11313147). In lane 1, the lysate was stored at -80C°C prior to Western Blotting.  The bands beneath the target band (40 kDa) are likey to be degradation products. In lane 3, the lysate was freshly made and used for Western Blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 37 seconds

All lanes:

Western blot - Anti-FOXF1 antibody [EPR27368-79] (ab308633) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HeLa 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: 39 kDa

Observed band size: 40 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-FOXF1 antibody [EPR27368-79] (AB308633)

All lanes:

Western blot - Anti-FOXF1 antibody [EPR27368-79] (ab308633) at 1/1000 dilution

All lanes:

Rat lung 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 1/20000 dilution

Predicted band size: 39 kDa

Observed band size: 40 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-FOXF1 antibody [EPR27368-79] (AB308633)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : colon (PMID : 11313147, 7957066), heart (PMID : 11313147, 7957066). The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29162563, 11313147). In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : 37 seconds

All lanes:

Western blot - Anti-FOXF1 antibody [EPR27368-79] (ab308633) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Mouse heart 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 1/20000 dilution

Predicted band size: 39 kDa

Observed band size: 40 kDa

false

Exposure time: 37s