8. 660 mM IAA prepared in 25 mM Ambic.
9. 100 mM Tris–HCl, pH 5.5.
10. Thermostatic bath.
Nanoparticles in Urine
Samples
1. Microfuge.
2. Ultrapure water.
3. Gold nanoparticles, AuNPs.
4. 2.5 M Tris–HCl, pH 8.8.
5. 500 mM fresh DTT.
6. 660 mM IAA prepared in 25 mM Ambic.
7. 1 M citric acid.
8. Citrate/citric acid buffer, pH 4.0 (100 mM citrate, 100 mM
citric acid).
9. Thermostatic bath.
2.3 SDS Gels
1. Microfuge.
2. Electrophoresis chambers.
3. Power Pac Universal Supply.
4. Ultrapure water.
5. SDS-PAGE loading buffer: 10% w/v SDS, 40 mM Tris-Base
(pH 6.8), 50% v/v glycerol, 0.1% v/v bromophenol blue, 10%
v/v β-mercaptoethanol.
6. Eppendorf Thermo Mixer
® .
7. 1.5 M Tris–HCl, pH 8.8. Store at 4
C.
8. 0.5 M Tris–HCl, pH 6.8. Store at 4
C.
9. Acrylamide/bis solution (29.2:0.8 acrylamide:bis). Store at
4
C in an amber bottle.
10. Gels were prepared as described in Note 3.
11. Ammonium persulfate: 10% solution in Milli-Q water.
12. N,N,N,N
0 -tetramethyl-ethylenediamine. Store at 4
C.
13. SDS-PAGE running buffer: 0.025 M Tris–HCl, pH 8.3,
0.192 M glycine, 0.1% SDS.
2.4 Colloidal
Coomassie Blue
Staining
1. Ultrapure water.
2. Methanol (MeOH).
3. Acetic acid (HCOOH).
4. Fixed/destaining solution: 40% (v/v) ethanol and 10% (v/v)
acetic acid.
5. Colloidal Coomassie Blue stain.
112
Maria del Pilar Chantada-Va ´ zquez et al.