Elettroforesi - PowerPoint PPT Presentation

Elettroforesi

(segue)

Pozzetti

Stacking gel

Running gel

(A) Analysis of binding of the b subunit cytoplasmic domain (bsol) to ECF1 and ECF1 (-). Mixtures of polypeptides were first analyzed by native agarose gel electrophoresis (A) through a 1% agarose gel.

Lanes from left to right:

I, ECF1 (-)

II, bsol ; III, 

IV, bsol +  ;

V, bsol + ( 1-134);

VI, bsol + ECF1 (-);

VII,  + ECF1 (-);

VIII, bsol +  + ECF1 (-);

IX, ECF1 (-) + ( 1-134);

X, bsol + ECF1 (-) + ( 1-134).

(B) Bands were excised from the agarose gel and separated on a 10-18% gradient of polyacrylamide in SDS.

Rodgers et al (1997) J. Biol. Chem. 272: 31058

• Coomassie brilliant blue R-250
• Coomassie brilliant blue G-250
• Silver stain
• Coloranti fluorescenti
• Trasferimento su membrana (blotting)

R-250

G-250

Riduzione di Ag+

Comparison of the sensitivity achieved with SYPRO, silver and Coomassie brilliant blue stains. Identical SDS-polyacrylamide gels were stained with A) SYPRO Orange protein gel stain, B) SYPRO Red protein gel stain; C) silver stain and D) Coomassie brilliant blue stain, according to standard protocols.

Tank

Semi-dry

Luminol

1 2 3 4 5 6

Lane 1: standard pei molecolari

Lane 2-6: campioni

-

Si

O

+

-

Effetto elettroendoosmotico