ATG12 Antibody (JE30-61)

Western Blot: ATG12 Antibody (JE30-61) [NBP3-32056]

Western Blot: ATG12 Antibody (JE30-61) [NBP3-32056] - Western blot analysis of ATG12 on different lysates with Rabbit anti-ATG12 antibody (NBP3-32056) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HCT 116 cell lysate
Lane 3: HEK-293 cell lysate
Lane 4: SH-SY5Y cell lysate

Lysates/proteins at 20 ug/Lane.

Predicted band size: 15 kDa
Observed band size: 55/20 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32056) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/ Immunofluorescence: ATG12 Antibody (JE30-61) [NBP3-32056]

Immunocytochemistry/ Immunofluorescence: ATG12 Antibody (JE30-61) [NBP3-32056] - Immunocytochemistry analysis of HCT 116 cells labeling ATG12 with Rabbit anti-ATG12 antibody (NBP3-32056) at 1/250 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 in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATG12 antibody (NBP3-32056) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: ATG12 Antibody (JE30-61) [NBP3-32056]

Flow Cytometry: ATG12 Antibody (JE30-61) [NBP3-32056] - Flow cytometric analysis of HCT 116 cells labeling ATG12.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32056, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).