Materials and Reagents

1. Plant tissues
   
2. Sodium acetate (Bioshop, catalog number: SAA305 )
   
3. EDTA (Bioshop, catalog number: EDT001 )
   
4. Dithiothreitol (DTT) (Bioshop, catalog number: DTT 002 )
   
5. Phenylmethyl sulfonyl fluoride (PMSF) (G-Biosciences, catalog number: 786-0555 )
   
6. Thiourea (Sigma-Aldrich, catalog number: T-7875 )
   
7. Polyvinyl (polypyrrodlidone) (PVPP) (Sigma-Aldrich, catalog number: P-6755 )
   
8. β-nicotinamide adenine dinucleotide reduced form (NADH) (Bioshop, catalog number: NAD002 )
   
9. Phosphoelnolpyryvate (PEP) (Biovectra, catalog number: 2552 )
   
10. MgCl₂ ((Biolynx, catalog number: 18641 )
    
11. Rabbit muscle lactate dehydrogenase (LDH) (Sigma-Aldrich, catalog number: L-2500 )
    
12. para-nitrophenyl-phosphate (pNPP, phosphatase substrate) (Sigma-Aldrich, catalog number: P-4744 )
    
13. Coomassie Brilliant blue G-250 (Serva, catalog number: 35050 C.!.42655 )
    
14. Bovine gamma globulin (BGG) (2.0 mg/ml) (Thermo Fisher Scientific, catalog number: 23212 )
    
15. NaOH
    
16. Extraction buffer (EB) (see Recipes)
    
17. Bradford working solution (see Recipes)
    
18. Acid phosphatase enzyme assay mix #1 (see Recipes)
    
19. Acid phosphatase enzyme assay mix #2 (see Recipes)
    
20. Bradford assay stock (see Recipes)
    

Equipment

1. Whatman #1 filter paper
   
2. 1.5 ml microfuge tubes
   
3. 10 and 25 μl Hamilton syringes
   
4. Pipetor
   
5. Small mortar and pestle
   
6. Eppendorf microfuge
   
7. 96 well polystyrene microtitre plate (flat bottom)
   
8. A computer supported microplate spectrophotometer (Spectromax Plus, Molecular Devices, Sunnyvale)
   

Procedure