GPX4 Antibody (YA399)

Western blot analysis of extracts from HEK293 (lane 2(40μg) and HEK293 (lane 3(20μg)using GPX4 (HY-P80450) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% defatted milk powder in TBST for 2 hour at room temperature. The primary antibody (HY-P80450, 1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% defatted milk powder in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

Western blot analysis of extracts from HT-1080 (lane 1(20μg)) 、HepG2 (lane 2(20μg)) 、A549 (lane 3(20μg)) 、HEK293 (lane 4(20μg)) 、HCT116 (lane 5(20μg)) 、Jurkat (lane 6(20μg)) 、U937 (lane 7(20μg)) and Raw 264.7 (lane 8(20μg)) using GPX4 Antibody (HY-P80450) . Proteins were transferred to a PVDF membrane and blocked with 5% nonfat dry milk in TBST for 1.5 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Hsp90, 1/10000) was used in 5% nonfat dry milk in TBST at 4℃ overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

HT-1080 cells were seeded at 60000 cells/well in a 24 well plate and were cultured overnight. Then, cells were treated with Erastin (HY-15763) (0-100 nM) for 24 h, and the expression of GPX-4 was measured. The results indicated that Erastin inhibits GPX-4 expression. Primary antibody: GPX-4 antibody (HY-P80450), GAPDH antibody (HY-P80137).

HT-1080 cells were seeded at 60000 cells/well in a 24 well plate and were cultured overnight. Then, cells were treated with RSL3 (HY-100218A) (0-20 nM) for 24 h, and the expression of GPX-4 was measured. The results indicated that RSL3 inhibits GPX-4 expression. Primary antibody: GPX-4 antibody (HY-P80450), GAPDH antibody (HY-P80137).

Immunohistochemical analysis of paraffin-embedded mouse testis tissue using GPX4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80450, 1:1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse testis tissue using GPX4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80450, 1:1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue using GPX4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80450, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue using GPX4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80450, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Flow cytometric analysis of 1X10^6 HEK293 cells labeling GPX4 Antibody (HY-P80450, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1μg/mL dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

Immunocytochemistry analysis of 293 cells labeling GPX4 with GPX4 Antibody (HY-P80450) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GPX4 Antibody (HY-P80450) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Immunocytochemistry analysis of C6 cells labeling GPX4 with GPX4 Antibody (HY-P80450) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GPX4 Antibody (HY-P80450)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Immunocytochemistry analysis of Hela cells labeling GPX4 with GPX4Antibody (HY-P80450) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GPX4 Antibody (HY-P80450) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Immunocytochemistry analysis of HepG2 cells labeling GPX4 with GPX4 Antibody (HY-P80450) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GPX4 Antibody (HY-P80450)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).