- GTX22792 - GeneTex P4HB antibody

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Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Western blot analysis of PDI in 25 ug of HepG2, HeLa and NIH-3T3 cell lysates. Proteins were transferred to a PVDF membrane and blocked at 4¢XC overnight. The membrane was probed with PDI antibody [RL90] at a dilution of 1:1000 overnight at 4¢XC, washed in TBST, and probed with an HRP-conjugated secondary antibody. Chemiluminescent detection was performed.

Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Western blot analysis of PDI was performed by loading 25 ug of HepG2 (Lane 1), Hela (Lane 2) and NIH-3T3 (Lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a PDI monoclonal antibody (GTX22792) at a dilution of 1:1000 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. Results show a band at approx. 57 kDa.
Validation comment: WB

Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Immunofluorescent analysis of Phalloidin (blue) and PDI (green) in NIH 3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with PDI antibody [RL90] at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with a proper secondary antibody. Actin was stained with Dylight 350 Phalloidin and nuclei (red) were stained with DRAQ5. Images were taken at 20X magnification.

Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Flow cytometry analysis of PDI 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 and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with PDI antibody [RL90] at a dilution of 1 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using 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 PDI showing positive staining in the membrane and cytoplasm of NIH/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 and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a PDI monoclonal antibody (GTX22792) at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then 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 PDI showing positive staining in the membrane and cytoplasm 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 and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a PDI monoclonal antibody (GTX22792) at a dilution of 1 ug/test for 60 min at room temperature. Cells were then 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 PDI showing positive staining in the membrane and cytoplasm of K562 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a PDI monoclonal antibody (GTX22792) at a dilution of 1 ug/test for 60 min at room temperature. Cells were then 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

GTX22792