Materials and Reagents

1. Homogenized plant tissues
   
2. Liquid nitrogen
   
3. 30% acrylamide stock (29: 1 acrylamide: bisacrylamide) (Bio-Rad Laboratories, catalog number: 161-0156 )
   
4. TEMED (Bio-Rad Laboratories, catalog number: 161-0801 )
   
5. Ammonium persulfate (Sigma-Aldrich, catalog number: A3678 )
   
6. 2D Quant Kit (GE Healthcare, catalog number: 80-6483-56 )
   
7. Trizma^® base (Sigma-Aldrich, catalog number: T1503 )
   
8. Bromphenol blue (Sigma-Aldrich, catalog number: B0126 )
   
9. Glycerol
   
10. Triton X-100 (Sigma-Aldrich, catalog number: T8532 )
    
11. Na₂HPO₄ (Sigma-Aldrich, catalog number: S3264 )
    
12. NaH₂PO₄ (Sigma-Aldrich, catalog number: 71505 )
    
13. N,N-Dimethylformamide (Fluka, catalog number: 72438 )
    
14. Fast blue B salt (Sigma-Aldrich, catalog number: D9805 )
    
15. α-naphthyl palmitate (Sigma-Aldrich, catalog number: N9875 )
    Note: α-naphthyl palmitate catalog number: N9875 is no longer available in Sigma-Aldrich, but is available in Santa Cruz Biotechnology, catalog number: CAS 15806-43-6.
    
16. Extraction buffer (see Recipes)
    
17. 12% separating gel (see Recipes)
    
18. 4% stacking gel (see Recipes)
    
19. 2 x SDS sample buffer (see Recipes)
    
20. 1 M sodium phosphate buffer (see Recipes)
    
21. Developing solution (see Recipes)
    

Equipment

1. Mortar and pestle
   
2. Refrigerated centrifuge 5810R (Eppendorf, catalog number: 5810000.017 ) or similar, equipped with a rotor for 2 ml microcentrifuge tubes
   
3. Magnetic stirrer
   
4. 2 ml microcentrifuge tube
   
5. Tubular glass vials
   
6. Mini-PROTEAN^® Tetra Cell (Bio-Rad Laboratories, catalog number: 165-8000 )
   

Software

1. Quantity One software (Bio-Rad Laboratories)

Procedure