Immunofluorescence Staining of Multidrug Resistance-Associated Protein (MRP-2)



April 23, 2019

EP154 v1.0

Goals:

Key Steps:

Other Required Materials:

Visualize expression of multidrug resistance-associated protein (MRP-2) in fixed Emulate Organ-Chips

  • Immunofluorescence staining in Organ-Chips
  • Anti-MRP-2 antibody [M2 III-6] (Abcam, ab3373)
  • 4% paraformaldehyde (PFA)
  • Saponin
  • PBS
  • BSA
  • Normal goat serum (or other serum from the species the secondary antibody was raised in)
  • Alexa Fluor® 488-conjugated goat anti-mouse IgG secondary antibody (or other anti-mouse secondary antibody)
  • Fluorescence microscope

Method

Sample type

Fixed Organ-Chip

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

Fixative and incubation time (recommended)

4% PFA, 15 minutes at room temperature

Permeabilization and incubation time (recommended)

1% saponin in PBS, 30 minutes at room temperature

Blocking buffer and incubation time (recommended)

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

Primary antibody dilution and incubation time (recommended)

Dilute anti-MRP-2 antibody [M2 III-6] (Abcam, ab3373)

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

Antibody host

Mouse

Secondary antibody dilution and incubation time (recommended)

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

Representative image

Image of MRP-2 (green) / nuclei (blue) staining indicating expression and polarization of MRP2 by hepatocytes in the human Liver-Chip (top channel)

More information on vendor site

https://www.abcam.com/mrp2-antibody-m2-iii-6-ab3373.html

 


 

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