iFluor™ 647 Conjugated Goat anti-rabbit IgG polyclonal Antibody

Immunocytochemistry analysis of Hela cells labeling Vimentin with Rabbit anti-Vimentin antibody (ET1610-39) at 1/400 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 3% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Vimentin antibody (ET1610-39) at 1/400 dilution in 3% BSA overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 647, HA1123) 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.

Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling Synaptophysin (ET1606-56) and beta Ⅲ tubulin (M0805-8).

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Synaptophysin (ET1606-56, red) at 1/200 dilution and beta Ⅲ tubulin (M0805-8, green) at 1/200 dilution overnight at 4 ℃, washed with PBS.

iFluor™ 647 conjugate-Goat anti-Rabbit IgG (HA1123) and iFluor™ 488 conjugate-Goat anti-Mouse IgG (HA1125) were used as the secondary antibodies at 1/500 dilution. DAPI was used as nuclear counterstain.

Flow cytometric analysis of HeLa cells labeling beta Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1602-4, 1/1,000) (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™ 647 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1123) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).