Biotin N-hydroxysuccinimide ester → Biotin

HRID 344635Notes

Reaction equation

HRID 344635 reaction equation
C1C(=O)N([O:8][C:9]([CH2:11][CH2:12][CH2:13][CH2:14][C@@H:15]2[S:19][CH2:18][C@@H:17]3[NH:20][C:21]([NH:23][C@H:16]23)=[O:22])=[O:10])C(=O)C1>CN(C=O)C.C([O-])(O)=O.[Na+]>[OH:10][C:9]([CH2:11][CH2:12][CH2:13][CH2:14][C@H:15]1[C@@H:16]2[C@@H:17]([NH:20][C:21]([NH:23]2)=[O:22])[CH2:18][S:19]1)=[O:8]

Participants

4

Reactant 1

HCID 6710714Biotin N-hydroxysuccinimide ester4×

Product 1

HCID 171548Biotin

Reagents, catalysts & solvents 2

HCID 6228DimethylformamideSolventHCID 516892Sodium BicarbonateSolvent

Procedure

Histone H1 was biotinylated to different degrees with increasing amounts of BHSE. 5 μg BHSE (dissolved in 10 μl DMF), 25 μg BHSE (in 10 μl DMF) and 250 μg BHSE in 10μl DMF were added to three solutions containing the same amount of histone H1 (1 mg in 300 μl of 50 mM NaHCO3) and incubated for 1 hour at 20° C. After dialysis against 5 mM sodium phosphate (pH 6.8) the three samples are stored at -20° C. The concentrations of the biotin-labelled histone H1 solutions are determined by comparison with unmodified histone H1 by SDS-polyacrylamide gel electrophoresis. The number of biotin residues in the three preparations was estimated by the tritium marker contained in BHSE and was found to be between 2.7 and 20 per histone H1 molecule.

Workup

  1. customAfter dialysis against 5 mM sodium phosphate (pH 6.8) the three samples
  2. customare stored at -20° C

Sources & evidence

ORD record
ord-5bf0f635159d482cb89e9366520da456
Source dataset
uspto-grants-1991_10
Patent number
US05053326

Reaction notes

Loading…