Aconitase 2 Antibody (YA2151)

MedChemExpress Validation
Western blot analysis of extracts from K562(lane 2(20ug) ,NIH/3T3(lane 3(20ug) and Neuro-2a(lane 4(20ug) using Aconitase 2 Antibody (HY-P82406) 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.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue using Aconitase 2 Antibody (YA2151). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) 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-P82406, 1/100) 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.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue using Aconitase 2 Antibody (YA2151). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) 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-P82406, 1/100) 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.

MedChemExpress Validation
Immunocytochemistry analysis of NIH/3T3 cells labeling Aconitase 2 Antibody (HY-P82406) 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 BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with Aconitase 2 Antibody (HY-P82406) at 1/50 dilution in BSA for Immunol Staining at 4 ℃ Overnight. AF488-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).

MedChemExpress Validation
Immunocytochemistry analysis of NIH/3T3 cells labeling Aconitase 2 Antibody (HY-P82406) at 1/200 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 BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with Aconitase 2 Antibody (HY-P82406) at 1/200 dilution in BSA for Immunol Staining at 4 ℃ Overnight. AF488-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).