Materials and Reagents

1. Species  
   Three Chlorella species were used: C. vulgaris, isolated from soil samples of Livingston Island, the South Shetland Archipelago, Antarctic; C. vulgaris strain 8/1, isolated in 1968 from thermal springs in the region of Rupite, Bulgaria, and cultivated in our laboratory since 1975 and Chlorella kesslery a mesophile, from the Trebon collection.
   Cultivation  
   Chlorella species were cultivated on TAP (Tris Acetate Phosphate) medium under continuous light of 60 μmol/m²/s and a temperature of 23 °C ± 0.1 °C in a Phytotron GC 400 growth chamber. The species were cultivated at this temperature because it is well known, that eurythermal algae, could be grown at a wide range of temperatures.
   
2. Rabbit polyclonal antibody HSP70B cytoplasmic (Agrisera, catalog number: AS06 175 )
   
3. Goat anti-rabbit IgG(H&l) HRP conjugated (Agrisera, catalog number: AS09 602 )
   
4. Coomassie brilliant blue G 250
   
5. Orthophosphoric acid (Valerus, catalog number: N 4420 )
   
6. Trichloroacetic acid (TCA)
   
7. Bovine serum albumin (BSA) (Applichem GmbH, catalog number: 1391 0025 )
   
8. Albumin fraction V (pH 7.0)
   
9. Medium Pure Nitrocellulose (NCM) (0.45 μm) (Bio-Rad Laboratories, catalog number: 162-0115 )
   
10. Filter paper
    
11. Sponge
    
12. 4CN (4-chloro-naphthol) (Bio-Rad Laboratories, catalog number: N170-6535 )
    
13. N,N′ N′ Tetramethylethylendiamine (TEMED) (Alfa Aesar, catalog number: N12536 )
    
14. Laemmli sample buffer (see Recipes)
    
15. Reagent of Bradford (see Recipes)
    
16. 5x Laemmli buffer (see Recipes)
    
17. Running buffer (see Recipes)
    
18. Transfer buffer (see Recipes)
    
19. SDS-PAGE gel (see Recipes)
    
20. 30% Acrylamide/N,N’-methylenebisacrylamide (AA/MBA) (see Recipes)  
    
21. 10% SDS (see Recipes)
    
22. 10% Ammonium Persulfate (see Recipes)
    
23. 1.5 M Tris HCl buffer (pH 8.8) (see Recipes)
    
24. 1.0 M Tris HCl buffer (pH 6.8) (see Recipes)
    
25. 4 M NaCl (see Recipes)
    
26. 1.0 M Tris HCl buffer (pH 7.5) (see Recipes)
    
27. 20% Tween 20 (see Recipes)
    
28. Blocking buffer (see Recipes)
    
29. Staining solution (see Recipes)
    
30. 5% CH₃COOH (see Recipes)
    
31. Washing solution (see Recipes)
    
32. 50 mM TBS-T buffer (see Recipes)
    
33. HRP color development solution (see Recipes)
    

Equipment

1. Motor
   
2. Silica  quartz sand 0.6 mm (Valerus, catalog number: N 1760 )
   
3. Centrifuge (Sigma-Aldrich, model: 1-15 K)
   
4. Electrophoresis chamber Transfer unit Hoefer miniVE electrophotesis and electrotransfer unit (Hoefer, model: SE300-10A-1.0 )
   
5. Mini Rocker Shaker MR-1
   

Software

1. Image J program

Procedure