CD130 Antibody (YA3990)

Western blot analysis of extracts from HUVEC (lane 2(20μg), HepG2 (lane 3(20μg), Raji (lane 4(20μg) and Hela (lane 5(20μg) using CD130 Antibody (HY-P84293) 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-P80993, 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 PC-12(lane 1(40μg) ),COS-7(lane 2(40μg) ),Hela(lane 3(40μg) )and HepG2(lane 4(40μg) ) using CD130 antibody(31685P). 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 HL-60 cells labeling CD130 Antibody(31685P,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 HUVEC cells labeling CD130 with CD130 antibody (HY-P84293) 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 CD130 antibody (HY-P84293) at 1/50 dilution in BSA for Immunol Staining at 4 ℃, Stay 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 HUVEC cells labeling CD130 with CD130 antibody (HY-P84293) at 1/100 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 CD130 antibody (HY-P84293) at 1/100 dilution in BSA for Immunol Staining at 4℃, Stay 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).

Immunofluorescence analysis of Hela cells labeling CD130 antibody (31685P) at 1/50 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 1% BSA in 10% negative goat serum for 1 hour at room temperature.Cells were then incubated with CD130 antibody (31685P) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Alexa Fluor® 488 Goat Anti-mouse IgG H&L (invitrogen A11001, green) was used as the secondary antibody at 1/500 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue)