All lanes : Anti-Glutathione Peroxidase 3/GPx-3 antibody [EPR22815-112] (ab256470) at 1/1000 dilution

Lane 1 : Mouse kidney lysate
Lane 2 : Mouse liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 25 kDa
Observed band size: 24/22 kDa why is the actual band size different from the predicted?


Exposure time: 48 seconds

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 20015939).

The 22-KDa band represents another Gpx3 isoform. 

Negative control: Mouse liver (PMID: 15294905).

Anti-Glutathione Peroxidase 3/GPx-3 antibody [EPR22815-112] (ab256470) at 1/1000 dilution (Purified) + Rat kidney lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 25 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?


Exposure time: 26 seconds

Blocking and dilution buffer: 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 20015939).

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat kidney tissue labeling Glutathione Peroxidase 3/GPx-3 with ab256470 at 1/100 (4.80 μg/ml) dilution followed by ab150077 AlexaFluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Positive staining on rat kidney (PMID: 21493731) is observed.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

 

Immunohistochemical analysis of 4% PFA fixed, 0.2% Triton X-100 permeabilized frozen mouse kidney tissue labeling Glutathione Peroxidase 3/GPx-3 with ab256470 at 1/100 (4.80 μg/ml) dilution followed by ab150077 AlexaFluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Positive staining on mouse kidney (PMID: 21493731) is observed.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling Glutathione Peroxidase 3/GPx-3 with ab256470 at 1/500 dilution (0.96µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat lung (PMID: 22355371) is observed. The section was incubated with ab256470 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Glutathione Peroxidase 3/GPx-3 with ab256470 at 1/500 dilution (0.96µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat kidney (PMID: 21493731, 27435875) is observed. The section was incubated with ab256470 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling Glutathione Peroxidase 3/GPx-3 with ab256470 at 1/500 dilution (0.96µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse lung (PMID: 22355371) is observed. The section was incubated with ab256470 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Glutathione Peroxidase 3/GPx-3 with ab256470 at 1/500 dilution (0.96µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse kidney (PMID: 21493731, 27435875) is observed. The section was incubated with ab256470 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).