- GTX12412 - GeneTex PPARA antibody

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Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Western blot analysis of PPAR alpha in 25 ug of HeLa, Jurkat and NIH-3T3 cell lysates. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4oC overnight. The membrane was probed with PPAR alpha antibody [3B6/PPAR] at a dilution of 1:1000 (HeLa and Jurkat) and 1:500 overnight at 4¢XC, washed in TBST, and probed with a proper secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed.

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Western blot analysis of PPAR alpha was performed by loading 25 ug of Hela (Lane 1), Jurkat (Lane 2), and NIH-3T3 cell lysates (Lane 3) and a molecular weight protein ladder onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4ºC overnight. The membrane was probed with a PPAR alpha monoclonal antibody (GTX12412) at a dilution of 1:1000 (Hela and Jurkat) and 1:500 (NIH-3T3) overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using ECL substrate. Results show a band at 52 kDa in all three cell lines.
Validation comment: WB

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Flow cytometry analysis of PPAR alpha in Hela cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde, washed with PBS, and incubated with PPAR alpha antibody [3B6/PPAR] at a dilution of 0.25 ug/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark with a proper secondary antibody, and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Flow cytometry analysis of PPAR alpha showing positive staining in the nucleus of 3T3 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde, washed with PBS, and incubated with PPAR alpha monoclonal antibody (GTX12412) at a dilution of 0.25 ug/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.
Validation comment: FACS

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Flow cytometry analysis of PPAR alpha showing positive staining in the nucleus of Hela cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde, washed with PBS, and incubated with PPAR alpha monoclonal antibody (GTX12412) at a dilution of 0.25 ug/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.
Validation comment: FACS

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Flow cytometry analysis of PPAR alpha showing positive staining in the nucleus of Jurkat cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde, washed with PBS, and incubated with PPAR alpha monoclonal antibody (GTX12412) at a dilution of 0.25 ug/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.
Validation comment: FACS

GTX12412