iFluor™ 488 Conjugated Actin Recombinant Rabbit Monoclonal Antibody [JJ09-29]

Immunocytochemistry analysis of NIH/3T3 cells labeling Actin with Rabbit anti-Actin antibody (HA720160F) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% normal goat serum for 1 hour at 37 ℃. Cells were then incubated with Rabbit anti-Actin antibody (HA720160F) at 1/100 dilution in 2% normal goat serum overnight at 4 ℃. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) were used as the secondary antibody at 1/800 dilution.

Immunofluorescence analysis of paraffin-embedded human colon carcinoma tissue labeling Actin (HA720160F).

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 antibody Actin (HA720160F, iFluor™ 488) at 1/50 dilution overnight at 4 ℃, washed with PBS. DAPI was used as nuclear counterstain.

Flow cytometric analysis of NIH/3T3 cells labeling Actin.

Cells were fixed and permeabilized. Then incubated for 30 minutes at +4℃ with Actin (HA720160F, red, 1ug/ml) and Rabbit IgG Isotype Control (iFluor™ 488, green, 1ug/ml). Unlabelled sample was used as a control (cells without incubation with primary antibody; black).