Cyclin D1/CCND1 Antibody (YA485)

Western blot analysis of extracts from C6 (lane 2(20μg), C6 (lane 3(40μg), NIH/3T3 (lane 4(20μg), NIH/3T3 (lane 5(40μg) using Cyclin D1 Antibody (HY-P80098) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80098, 1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature

Western blot analysis of extracts from Neuro-2a (lane 2(40μg),NIH/3T3 (lane 3(40μg), C6 (lane 4(40μg)using Cyclin D1/CCND1 Antibody (YA485). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

Western blot analysis of extracts from MCF7 (lane 1(20μg)) 、HepG2 (lane 2(20μg)) 、NIH-3T3 (lane 3(20μg)) and C6 (lane 4(20μg)) using Cyclin D1/CCND1 Antibody (HY-P80098) . Proteins were transferred to a PVDF membrane and blocked with 5% nonfat dry milk in TBST for 1.5 hour at room temperature. The primary antibody (1/2000) and Loading control antibody (β Tubulin, 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 WT Hela cells (lane 1(20μg)) and KD Hela cells (lane 2(20μg)) using Cyclin D1/CCND1 Antibody (HY-P80098) . Proteins were transferred to a PVDF membrane and blocked with 5% nonfat dry milk in TBST for 1.5 hour at room temperature. The primary antibody (1/2000) 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.

TEST

Immunocytochemistry analysis of NIH3T3 cells labeling Cyclin D1 with Cyclin D1 Antibody (HY-P80098) 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 Cyclin D1 Antibody (HY-P80098) 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 Neuro-2a cells labeling Cyclin D1 with Cyclin D1 Antibody (HY-P80098) 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 Cyclin D1 Antibody (HY-P80098)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).