GAPDH Antibody (YA418)

Western blot analysis of extracts from MCF-7(lane 2(20ug) , THP-1(lane 3(20ug)and HepG2(lane 4(20ug) using GAPDH Antibody (HY-P80137) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P83730, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) 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).

HCT-116 cells were seeded at 4*10⁵ cells/well in a 6-well plate and were cultured overnight. Then, cells are treated with 10 nM Actinomycin D (HY-17559) for 0-8 hours to assess the expression of MDM2 and p53 via Western blotting. The results demonstrated that Actinomycin D upregulated the expression of MDM2 and p53 in a time-dependent manner. Primary antibody: MDM2 antibody (HY-P83705), p53 antibody (HY-P80938), GAPDH antibody (HY-P80137).

Immunohistochemical analysis of paraffin-embedded rat liver tissue using GAPDH Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 at 1/100 dilution 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 rat liver tissue using GAPDH Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 at 1/100 dilution 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 Hela cells labeling GAPDH Antibody (YA418) (HY-P80137, red). Cells were stained with the primary antibody at 1/100 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 Hela cells labeling GAPDH with GAPDH Antibody (HY-P80137) at 1:50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with GAPDH Antibody (HY-P80137) at 1:50 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated 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 NIH-3T3 cells labeling GAPDH with GAPDH Antibody (HY-P80137) at 1:50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with GAPDH Antibody (HY-P80137) at 1:50 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated 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).