Tris(hydroxymethyl)aminomethane → Phospho-L-tyrosine

HRID 1826099

Reaction equation

HRID 1826099 reaction equation
CCCCCC[CH2:7][CH2:8][CH2:9][CH2:10][CH2:11][CH2:12][O:13]S([O-])(=O)=O.[Na+].C(O)[C:20]([NH2:25])([CH2:23]O)[CH2:21][OH:22].C1C(Cl)=CC(SC2C=C(Cl)C=CC=2[OH:42])=C(O)C=1.C1C(CC(N[P:56]([OH:59])([OH:58])=[O:57])C(O)=O)=CC=C(O)C=1>[O-][V]([O-])([O-])=O.[Na+].[Na+].[Na+]>[P:56]([O:13][C:12]1[CH:7]=[CH:8][C:9]([CH2:23][C@@H:20]([C:21]([OH:22])=[O:42])[NH2:25])=[CH:10][CH:11]=1)([OH:59])([OH:58])=[O:57]

Participants

6

Reactant 4

HCID 6503Tris(hydroxymethyl)aminomethaneHCID 7329FenticlorHCID 3423265Sodium Lauryl Sulfate2×HCID 11196273Anti-phosphotyrosine

Product 1

HCID 30819Phospho-L-tyrosine

Reagents, catalysts & solvents 1

HCID 61671Sodium orthovanadateCatalyst

Procedure

Cells were starved (24 hours for MCF-7, 4 hours for 32D derived cell lines) before addition of the indicated amounts of growth factors. Total cell lysates were prepared by addition of SDS PAGE sample buffer (1% SDS, 0.15M Tris pH 8.6, 5% BME and 1 mM Sodium Ortho Vanadate) directly to cells. Cell lysates and were run on 8–16% Tris-Glycine gradient gels (Novex). Proteins were transferred onto Hybond ECL nitrocellulose membranes (Amersham) and were immunoblotted with anti-phosphotyrosine MAb.

Workup

  1. additionbefore addition of the indicated amounts of growth factors

Sources & evidence

ORD record
ord-2a606c8a2cd546bfa6dd419a50819e9c
Source dataset
uspto-grants-2006_05
Patent number
US07049409B2

Linked notes

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