Sucrose → Tris Tricine

HRID 1618866

Reaktionsgleichung

Reaktionsgleichung von HRID 1618866
[NH:1]([C:6]([CH2:11][OH:12])([CH2:9][OH:10])[CH2:7][OH:8])[CH2:2][C:3]([OH:5])=[O:4].CCCCCCCCCCCCOS([O-])(=O)=O.[Na+].C(O)[C@H]1O[C@H](O[C@]2(CO)O[C@H](CO)[C@@H](O)[C@@H]2O)[C@H](O)[C@@H](O)[C@@H]1O.C1C=CC2S(=O)(=O)OC(C3C=C(Br)C(O)=C(Br)C=3)(C3C=C(Br)C(O)=C(Br)C=3)C=2C=1.CC[C@@H]([C@@H]1NC(=O)[C@@H](CCCN)NC(=O)[C@@H](NC([C@@H](NC([C@H](NC([C@@H](NC([C@H]2N=C([C@@H](N)[C@H](CC)C)SC2)=O)CC(C)C)=O)CCC(O)=O)=O)[C@H](CC)C)=O)CCCCNC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](CC(O)=O)NC(=O)[C@H](CC2NC=NC=2)NC(=O)[C@@H](CC2C=CC=CC=2)NC1=O)C>C(O)C(N)(CO)CO.Cl.CO.C(O)(=O)C>[CH2:7]([OH:8])[C:6]([NH2:1])([CH2:11][OH:12])[CH2:9][OH:10].[NH:1]([C:6]([CH2:9][OH:10])([CH2:11][OH:12])[CH2:7][OH:8])[CH2:2][C:3]([OH:5])=[O:4]

Komponenten

9

Edukt 5

HCID 5988SucroseHCID 8272Bromophenol BlueHCID 79784TricineHCID 3423265Sodium Lauryl Sulfate2×HCID 109094301405-87-4

Produkt 1

HCID 86608588Tris Tricine

Reagenzien, Katalysatoren, Lösungsmittel 3

HCID 176Acetic AcidLösungsmittelHCID 887MethanolLösungsmittel2×HCID 93573Tromethamine hydrochlorideLösungsmittel3×

Versuchsvorschrift

Tris tricine SDS-PAGE electrophoresis was performed according to Schagger and von Sagow [1987] using precast 16.5% T gels (Biorad, Hercules, Calif.). The anode buffer consisted of 0.2M Tris-HCl, pH 8.9 and the cathode buffer consisted of 0.1M Tris-HCl, 0.1M Tricine, 0.1% SDS, pH 8.25. Samples were diluted in 10 μl of 50 mM Tris-HCl, 4% (w/v) SDS 12% (w/v) sucrose, 5% (v/v) β morcaptoethanol, and trace of bromophenol blue, pH 6.8. After denaturation at 95° C. for 5 min, samples were loaded onto the gel. Gels were run at 80V for 3 hours. After electrophoresis, gels were fixed in 40% methanol, 10% acetic acid for 30 min. Gels were then stained with Colloidal blue coomassie G250 overnight and destained in 30% methanol. Bands to be identified were immediately cut, placed in an eppendorf and kept at 4° C. until further analysis. The apparent molecular masses were determined by running polypeptide molecular weight (MW) standards: Triosephophate isomerase MW 26,625; Myoglobin MW 16,950; α-lactalbumin MW 14,437; Aprotinin MW 6,512; Insulin b chain, oxidised MW 3,496 and Bacitracin MW 1,423 (Biorad).

Aufarbeitung

  1. waitGels were then stained with Colloidal blue coomassie G250 overnight
  2. customkept at 4° C. until further analysis

Quellen und Nachweise

ORD-Eintrag
ord-bfbb460e2fb24ba29a1e7521fe1ff3da
Quelldatensatz
uspto-grants-2014_07
Patentnummer
US08772042B2

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