Lactate Dehydrogenase Antibody (YA316)

MedChemExpress Validation
Western blot analysis was performed on protein extracts (25 μg) from 293T (lane 2), A549 (lane 3), NIH/3T3 (lane 4), MCF-7 (lane 5), U87 (lane 6) and HepG2 (lane 7) using Lactate Dehydrogenase antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot^® Turbo^™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Myocardium tissue using Lactate Dehydrogenase antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80422, 1:10000 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Tonsil tissue using Lactate Dehydrogenase antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80422, 1:10000 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Tonsil tissue using Lactate Dehydrogenase antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80422, 1:10000 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Tonsil tissue using Lactate Dehydrogenase antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80422, 1:10000 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Ovarian Cancer‌ tissue using Lactate Dehydrogenase antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80422, 1:10000 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Kidney cancer tissue using Lactate Dehydrogenase antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80422, 1:10000 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded Human Glioma tissue using Lactate Dehydrogenase Antibody(HY-P80422, 1/1000) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded Human Endometrial cancer tissue using Lactate Dehydrogenase Antibody(HY-P80422, 1/1000) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded Human pancreas tissue using Lactate Dehydrogenase Antibody(HY-P80422, 1/1000) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

MedChemExpress Validation
Flow cytometric analysis of 1X10⁶ HeLa cells labeling Lactate Dehydrogenase Antibody (HY-P80422, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. AF488-conjugated 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).

MedChemExpress Validation
Immunocytochemistry analysis of HeLa cells labeling Lactate Dehydrogenase with Lactate Dehydrogenase Antibody (HY-P80422) 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 Lactate Dehydrogenase Antibody (HY-P80422) 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).

MedChemExpress Validation
Immunocytochemistry analysis of HeLa cells labeling Lactate Dehydrogenase with Lactate Dehydrogenase Antibody (HY-P80422) at 1/100 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 Lactate Dehydrogenase Antibody (HY-P80422) at 1/100 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).