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Home > Proteomics Services > 2D Western Blot: From Samples to Western Blot
2D Western Blot: From Samples To Western Blot
More Examples on 2D Western Blot

Tissues / cells
Tissues or cells
Protein extraction
Protein extraction
Protein assay
Protein labeling

2D Gel or 2D DIGE gel
2D Gel or 2D DIGE Gel
2D Gel Image (1 sample)

2D Gel Image
2D DIGE Gel Image (2 samples)

2D DIGE gel image
Image captured from the gel (no staining)

Gel transfer to Membrane
 
2D Membrane Image (1 sample)

2d membrance
2D DIGE Membrane Image (2 samples)

2d dige membrane
Image captured from 2D membrane (no staining)
 
Fluorescent 2D Western Blot
  

2D Western Blot: one color

2d western blot

2D Western Blot: two color

2d western blot
2D Western Blot
 
Spot matching

2D Western Blot: Spot matching

Stripping and re-blotting with other antibodies
Stripping and re-blotting

   
Protein ID by Mass Spectrometry
Protein ID by Mass Spectrometry
   
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2D Western blot:

Mouse liver samples were prepared using 2D lysis buffer. 100 ug of proteins was labeled and used for 2D electrophoresis or 2D DIGE using pH 4-7 linear IPG strip according to Applied Biomics protocol. Left: standard 2D gel; Right: 2D DIGE gel. Following 2D electrophoresis, the images were scanned using Typhoon 9400; then the 2D gel was subject to transfer to membrane; after protein transfer, the 2D membrane images were scanned again; and followed by double-color Western blot (left) to reveal keratin 8 (red) and keratin 18 (green) including their modified isoforms by CF555-labeled goat anti-rabbit Ig and CF647-labeled goat anti-mouse Ig (Biotium, Hayward, California); after the scanning, the 2D membrane was stripped and re-blocked followed by Western blot to reveal beta-actin. For 2D DIGE gel (right panels) membrane, using mouse anti-human keratin 8 to reveal all the K8 and its posttranslational modified forms (under the disease condition); secondary antibody is CF488-labeled goat anti-mouse IgG from Biotium.
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