Choline Acetyltransferase Recombinant Rabbit Monoclonal Antibody [JA67-11]

Western blot analysis of Choline Acetyltransferase on different lysates with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16) at 1/5,000 dilution.

Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
Lane 2: Neuro-2a cell lysate (negative) (20 µg/Lane)
Lane 3: Mouse brain tissue lysate (40 µg/Lane)
Lane 4: Mouse cerebellum tissue lysate (40 µg/Lane)
Lane 5: Rat brain tissue lysate (40 µg/Lane)
Lane 6: Rat cerebellum tissue lysate (40 µg/Lane)

Predicted band size: 83 kDa
Observed band size: 50/75 kDa

Exposure time: 5 seconds; ECL: K1801;

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 (ET1704-16) at 1/5,000 dilution was used in primary antibody dilution (K1803) at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of HeLa cells labeling Choline Acetyltransferase with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16) 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 in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of HeLa cells labeling Choline Acetyltransferase.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1704-16, 1μg/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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).