Wnt-1 Antibody (YA3644)

Western blot analysis of extracts from NIH3T3 (lane 2(20μg), 3T3-L1 (lane 3(20μg) and Hela (lane 4(20μg) using WNT1 (20378) Mouse 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 IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.

Western blot analysis of extracts from NIH/3T3(lane 1(40μg) ) and 3T3-L1(lane 2(40μg) ) using WNT1 antibody(20378P). Proteins were transferred to a NC membrane and blocked with 5% Skim milk in TBST for 2 hour at room temperature. The primary antibody ( 1/1000) and Loading control antibody (GAPDH，20035, 1/1000) was used in 5% Skim milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

Flow cytometric analysis of 1X10^6 Hela cells labeling WNT1 Antibody(20378P,green). Cells were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Then stained with the primary antibody at 1/400 dilution overnight at 4℃.Alexa Fluor® 488 Goat Anti-mouse IgG H&L (invitrogen A11001) was used as the secondary antibody at 1/1,000 dilution for 45 minutes at room temperature. Mouse IgG Isotype Control (Invitrogen 14-4714-B2, gray) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (red).

Immunocytochemistry analysis of HeLa cells labeling INT1 Antibody (HY-P83947) 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 INT1 Antibody (HY-P83947) at 1/50 dilution in BSA for Immunol Staining at 4 ℃overnight. AF488-conjugated AffiniPure 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 mouse 3T3 cells labeling INT1 Antibody (HY-P83947) 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 INT1 Antibody (HY-P83947) at 1/50 dilution in BSA for Immunol Staining at 4 ℃overnight. AF488-conjugated AffiniPure 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).