beta III Tubulin Antibody (YA586)

Western blot analysis was performed on extracts from U-87MG (lane 1, 15 μg), 3T3 (lane 2, 15 μg), C6 (lane 3, 15 μg), Mouse brain (lane 4, 15 μg), and Rat brain (lane 5, 15 μg) using beta III Tubulin Rabbit mAb.Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST at 4°C overnight.The primary antibody (1:5000 dilution) and the loading control antibody (GAPDH, HY-P80137, 1:20000 dilution) were incubated in 5% non-fat milk in TBST for 1 hour at 37°C.Goat Anti-Rabbit IgG-HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

Western blot analysis of extracts from WT Hela cells (lane 1(20μg)) and KD Hela cells (lane 2(20μg)) using beta III Tubulin Antibody (HY-P80570) . Proteins were transferred to a PVDF membrane and blocked with 5% BSA 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.

Western blot analysis of extracts from NIH/3T3(lane 2(20μg) and NIH/3T3(lane 3(40ug)using beta III Tubulin Antibody (HY-P80570) 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-P80438, 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.

Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using beta III Tubulin 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 (HY-P80570, 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.

Immunocytochemistry analysis of SH-SY5Y cells labeling beta III Tubulin with beta III Tubulin Antibody (HY-P80570) 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 beta III Tubulin Antibody (HY-P80570) 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 U87 cells labeling beta III Tubulin with beta III Tubulin Antibody (HY-P80570) 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 beta III Tubulin Antibody (HY-P80570) 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).