Cytokeratin 19 Antibody

Western blot analysis of extracts from A549(lane 2(20ug) , MCF-7(lane 3(20ug) and HepG2(lane 4(20ug),K562(lane 5(20ug) using Cytokeratin 19 Antibody (HY-P81183) 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.

Flow Cytometry analysis of MCF-7 cells labelling Cytokeratin 19(red) with Cytokeratin 19 Antibody (anti-Cytokeratin 19) (HY-P81183). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then cells were stained with the primary antibody at 1/200 dilution for an hour at 4℃. Goat Anti-Rabbit IgG H&L (AF488) (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control(HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

Immunofluorescence analysis of HEC1A cells labeling Cytokeratin 19 with Cytokeratin 19 (HY-P81183) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA/10% normal goat serum for 1 hour at room temperature. Cells were then incubated with Cytokeratin 19 (HY-P81183) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Alexa Fluor® 647-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P80952) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Immunofluorescence analysis of MCF7 cells labeling Cytokeratin 19 with Cytokeratin 19 (HY-P81183) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA/10% normal goat serum for 1 hour at room temperature. Cells were then incubated with Cytokeratin 19 (HY-P81183) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Alexa Fluor® 647-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P80952) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Immunocytochemistry analysis of MCF-7 cells labeling Cytokeratin 19 with Cytokeratin 19 Antibody (HY-P81183) 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 Cytokeratin 19 Antibody (HY-P81183) at 1/50 dilution in BSA for Immunol Staining at 4 ℃,stay 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).

Immunocytochemistry analysis of HCT116 cells labeling Cytokeratin 19 with Cytokeratin 19 Antibody (HY-P81183) 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 Cytokeratin 19 Antibody (HY-P81183) at 1/50 dilution in BSA for Immunol Staining at 4 ℃,stay 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).