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Elettroforesi. (segue). SDS-PAGE. Pozzetti. Stacking gel. Running gel. Elettroforesi di proteine in condizioni native.

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sds page
SDS-PAGE

Pozzetti

Stacking gel

Running gel

elettroforesi di proteine in condizioni native
Elettroforesi di proteine in condizioni native

(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

visualizzazione delle proteine separate
Visualizzazione delle proteine separate
  • Coomassie brilliant blue R-250
  • Coomassie brilliant blue G-250
  • Silver stain
  • Coloranti fluorescenti
  • Trasferimento su membrana (blotting)
coomassie
Coomassie

R-250

G-250

sensibilit a confronto
Sensibilità a confronto

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.

western blot
Western blot

Tank

Semi-dry

risultato
Risultato

1 2 3 4 5 6

Lane 1: standard pei molecolari

Lane 2-6: campioni

elettroforesi capillare

-

Si

O

Elettroforesi capillare

+

-

Effetto elettroendoosmotico