NSE Antibody (YA1006)

Western blot analysis of extracts from HepG2(lane 2(20μg) , SH-SY5Y (lane 3(20μg) ,U-87MG(lane 4(20μg)and NIH/3T3( lane 5(20μg) using NSE Antibody (HY-P81290B). 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-P80993，1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (HY-P8004，1/10,000) was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human brain tissue using NSE Antibody (YA1006). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81290B, 1/100) 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 neutral balsam.

Immunohistochemical analysis of paraffin-embedded human glioma tissue using NSE Antibody (YA1006). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81290B, 1/100) 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 neutral balsam.

Immunohistochemical analysis of paraffin-embedded human brain cancer tissue using NSE Antibody (YA1006). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81290B, 1/100) 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 neutral balsam.

Tyramide signaling amplification based immunofluorescence analysis of paraffin-embedded human brain tissue using NSE Antibody (YA1006) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81290B, 1/500) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer and fluorescent tyramide signal amplification system.Immunostaining was performed with Vari Fluor 594 TSA (200×)（HY-D1835）. Tissues were counterstained with DAPI (blue) and mounted with Anti-fade fluorescence mounting medium.

Tyramide signaling amplification based immunofluorescence analysis of paraffin-embedded human glioma tissue using NSE Antibody (YA1006) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81290B, 1/500) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer and fluorescent tyramide signal amplification system.Immunostaining was performed with Vari Fluor 594 TSA (200×)（HY-D1835）. Tissues were counterstained with DAPI (blue) and mounted with Anti-fade fluorescence mounting medium.

Tyramide signaling amplification based immunofluorescence analysis of paraffin-embedded human brain cancer tissue using NSE Antibody (YA1006) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81290B, 1/500) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer and fluorescent tyramide signal amplification system.Immunostaining was performed with Vari Fluor 594 TSA (200×)（HY-D1835）. Tissues were counterstained with DAPI (blue) and mounted with Anti-fade fluorescence mounting medium.

Tyramide signaling amplification based immunofluorescence analysis of paraffin-embedded human small cell lung cancer tissue using NSE Antibody (YA1006). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81290B, 1/500) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer and fluorescent tyramide signal amplification system.Immunostaining was performed using a Bicolor signal amplification fluorescence staining kit. Tissues were counterstained with DAPI (blue) and mounted with Anti-fade fluorescence mounting medium.

Immunocytochemistry analysis of HeLa cells labeling NSE with NSE Antibody (HY-P81290B) 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 NSE Antibody (HY-P81290B) at 1/100 dilution in quick block buffer overnight at 4 ℃AF488-conjugated Goat Anti-Mouse IgG H&L（HY-P8005, 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 SH-SY5Y5 cells labeling NSE with NSE Antibody (HY-P81290B) 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 NSE Antibody (HY-P81290B) at 1/100 dilution in quick block buffer overnight at 4 ℃AF488-conjugated Goat Anti-Mouse IgG H&L（HY-P8005, 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).