Recombinant Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
The cells were treated with 0.25% trypsin (containing 0.02% EDTA) to produce a single cell suspension, followed by centrifugation at 1,000 × g for 5 min, and the supernatant was discarded. After washing in PBS, the cells were centrifuged and resuspended and then fixed with 95% ethanol.	Vimentin Ab 1:25; 40 µl at 4°C for 20 h)

Upstream tips
The cells were treated with 0.25% trypsin (containing 0.02% EDTA) to produce a single cell suspension, followed by centrifugation at 1,000 × g for 5 min, and the supernatant was discarded. After washing in PBS, the cells were centrifuged and resuspended and then fixed with 95% ethanol.
Protocol tips
Vimentin Ab 1:25; 40 µl at 4°C for 20 h)

Publication protocol

ADSCs of the third generation in a good growth state were selected along with cells from the control and ECM groups at 1, 3, 5, 7 and 9 days post-induction. The cells were treated with 0.25% trypsin (containing 0.02% EDTA) to produce a single cell suspension, followed by centrifugation at 1,000 × g for 5 min, and the supernatant was discarded. After washing in PBS, the cells were centrifuged and resuspended and then fixed with 95% ethanol. ADSCs of the third generation were mixed with fluorescein-isothiocyanate (FITC)-labeled monoclonal antibodies targeting CD29 (cat. no. ab24693; 1:200), CD31 (cat. no. ab9498; 1:20), CD40 (cat. no. ab224639, 1:600), CD44 (cat. no. ab189524; 1:40), CD45 (cat. no. ab25386, 1:200), CD90 (cat. no. ab212885; 1:500), CD105 (cat. no. ab44967; 1:200) and HLA-DR (cat. no. ab136320; 1:500). The typical marker of fibroblasts, vimentin, was used as the primary antibody for the induced cells (cat. no. ab92547; 1:25; 40 µl at 4°C for 20 h), and FITC-labeled goat anti-mouse IgG was used as the secondary antibody (cat. no. ab6785; 1:25; 40 µl at 4°C for 4 h). All antibodies were from Abcam (Cambridge, MA, USA). The expression of cell surface markers and the proportion of fibroblasts were measured by flow cytometry. There was one tube without any primary antibody that served as the negative control for each test. The experiment was repeated three times.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Flow cytometry Anti-bodies Human - Vimentin using Recombinant Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547) from Abcam.

Manufacturer protocol

Download the product protocol from Abcam for Recombinant Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Flow cytometry Anti-bodies Human - Vimentin using Recombinant Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547) from Abcam. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.