Immunofluorescence Staining of Vimentin

April 23, 2019

EP150 v1.0



Key Steps:

Other Required Materials:

Visualize vimentin via fluorescent imaging in fixed Emulate Organ-Chips

  • Immunofluorescence staining in Organ-Chips
  • Anti-vimentin antibody [SP20] (Abcam ab16700) 4% paraformaldehyde (PFA)
  • 10% Saponin
  • PBS
  • BSA
  • Normal goat serum (or other serum from the species the secondary antibody was raised in)
  • Alexa Fluor® 488-conjugated goat anti-rabbit IgG secondary antibody (or other anti-rabbit secondary)
  • Fluorescence microscope


Sample type

Fixed Organ-Chip

See Emulate Protocol EP137 Fixation and Immunofluorescence (IF) Staining.

Recommended fixative and incubation time

4% PFA, 15 mins at room temperature

Recommended permeabilization and incubation time

1% Saponin in PBS, 30 minutes at room temperature

Recommended blocking buffer and incubation time

1% BSA, 10% goat serum in PBS, overnight at 4°C

Recommended primary antibody dilution and incubation time

Anti-Vimentin [SP20] (Abcam ab16700)

1:1000 dilution in blocking buffer, overnight at 4°C

Recommended antibody host


Recommended secondary antibody dilution and incubation time

Alexa Fluor® 488-conjugated goat anti-rabbit IgG secondary antibody 1:500 dilution in blocking buffer, 2 hours at room temperature in the dark.

Representative image

Image of human stellate cells stained by vimentin (green) in the human Liver-Chip (bottom channel)

More information on vendor site



© Emulate, Inc., 2019. All rights reserved.

Emulate® and the Emulate logo are registered trademarks of Emulate, Inc.

Alexa Fluor® is a registered trademarks of Thermo Fisher Scientific.

The technology disclosed in this document may be covered by one or more patents or patent applications owned by or licensed to Emulate, Inc. No license is granted herein. You are solely responsible for determining whether you have all intellectual property rights that are necessary for your intended use of Emulate products or protocols, and whether you are required to obtain any additional intellectual property rights from a third party. Further information is available by contacting Emulate.