Anti-Vimentin antibody (ab137321)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	hPDLCs were rinsed with PBS for 3 times and fixed in 4% paraformaldehyde for 20 min. The fixed cells were treated with 0.5% Triton X-100 and 3% hydrogen peroxide for 15 min. The cells were then incubated for 1 h at 37 °C with rabbit anti-vimentin antibody (1:500).	Thereafter, the cells were washed with PBS for 3 times and incubated with corresponding secondary antibodies for 30 min at 37 °C. Finally, the cells were stained with a DAB kit (ZSGB-BIO, Beijing, China) and haematoxylin and viewed under an inverted microscope (Olympus, IX71, Tokyo, Japan).

Protocol tips
hPDLCs were rinsed with PBS for 3 times and fixed in 4% paraformaldehyde for 20 min. The fixed cells were treated with 0.5% Triton X-100 and 3% hydrogen peroxide for 15 min. The cells were then incubated for 1 h at 37 °C with rabbit anti-vimentin antibody (1:500).
Downstream tips
Thereafter, the cells were washed with PBS for 3 times and incubated with corresponding secondary antibodies for 30 min at 37 °C. Finally, the cells were stained with a DAB kit (ZSGB-BIO, Beijing, China) and haematoxylin and viewed under an inverted microscope (Olympus, IX71, Tokyo, Japan).

Publication protocol

The expression of keratin and vimentin in hPDLCs was then characterized by immunostaining. Briefly, hPDLCs were rinsed with PBS for 3 times and fixed in 4% paraformaldehyde for 20 min. The fixed cells were treated with 0.5% Triton X-100 and 3% hydrogen peroxide for 15 min. The cells were then incubated for 1 h at 37 °C with a primary mouse anti-pan-cytokeratin antibody (1:500) and a primary rabbit anti-vimentin antibody (1:500) (Abcam, Cambridge, MA, USA). Thereafter, the cells were washed with PBS for 3 times and incubated with corresponding secondary antibodies for 30 min at 37 °C. Finally, the cells were stained with a DAB kit (ZSGB-BIO, Beijing, China) and haematoxylin and viewed under an inverted microscope (Olympus, IX71, Tokyo, Japan).

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Manufacturer protocol

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