Immunofluorescence Staining of Bile Salt Export Pump (BSEP) Transporter



April 9, 2019

EP153 v1.0

 

Goals:

Key Steps:

Other Required Materials:

Visualize bile salt export pump (BSEP) transporter in hepatocytes in fixed Liver-Chip

  • Immunofluorescence staining in Liver-Chip
  • Anti-ABCB11 antibody (Abcam ab155421)
  • 4% paraformaldehyde (PFA)
  • 10% saponin
  • PBS
  • BSA
  • Normal goat serum (or other serum from the species the secondary antibody was raised in)
  • Alexa Fluor™ 555-conjugated goat anti-rabbit IgG secondary antibody (or other anti-rabbit secondary antibody)
  • Fluorescence microscope

Method

Sample type

Fixed Liver-Chip

See Protocol EP137 Fixation and Immunofluorescence (IF) Staining.

Recommended fixative and incubation time

4% PFA, 15 minutes 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-ABCB11 antibody (Abcam, ab155421)

1:100 dilution in blocking buffer, overnight at 4°C

Recommended antibody host

Rabbit

Recommended secondary antibody dilution and incubation time

Alexa Fluor™ 555-conjugated goat anti-rabbit IgG secondary antibody 1:500 in blocking buffer, 2 hours at room temperature in the dark

Representative image

Image of BSEP transporter (red) / nuclei (blue) staining indicating localization of BSEP transporter in the bile canalicular membrane of hepatocytes in the human Liver-Chip (top channel).

More information on vendor site

https://www.abcam.com/abcb11-antibody-ab155421.html

 


 

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