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Biotin-HPDP
Sulfhydryl-reactive biotinylation reagent, pyridyldithiol-activated

Catalog No.A8008
Size Price Stock Qty
10mM (in 1mL DMSO)
$50.00
In stock
10mg
$45.00
In stock
50mg
$180.00
In stock
100mg
$225.00
In stock

Tel: +1-832-696-8203

Email: [email protected]

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Sample solution is provided at 25 µL, 10mM.

Product Citations

1. Das S, Shklyaev OE, et al. "Harnessing catalytic pumps for directional delivery of microparticles in microchambers. "Nat Commun. 2017 Feb 17;8:14384. PMID:28211454
2. Ortiz-Rivera, Isamar, et al. "Convective flow reversal in self-powered enzyme micropumps." Proceedings of the National Academy of Sciences 113.10 (2016): 2585-2590. PMID:26903618
3. Sambeeta Das. "DESIGNS FOR DIRECTING MOTION AT THE MICRO-AND NANOSCALE." The Pennsylvania State University.August 2016.
4. Isamar Ortiz‐Rivera, Taylor M.Courtney,Ayusman Sen. "Enzyme Micropump‐Based Inhibitor Assays." Advanced Functional Materials.April 5, 2016.

Quality Control

Chemical structure

Biotin-HPDP

Related Biological Data

Biotin-HPDP

Related Biological Data

Biotin-HPDP

Related Biological Data

Biotin-HPDP

Related Biological Data

Biotin-HPDP

Biological Activity

Description Biotin-HPDP is a sulfhydryl-reactive biotinylation agent.
Targets            
IC50            

Protocol

Biotinylation method [1]:

Sample

S-nitrosylated proteins

Preparation method

Soluble in DMSO or DMF.

Reaction Conditions

50mM, 25 ℃ for 1 h

Applications

Prepare biotin-HPDP as a 50mM suspension in DMSO, Dilute with DMF to a final concentration of 4 mM. Add 1:3 volume of Labeling Solution and 1:50 volume of Ascorbate Solution(50mM) to the blocked protein samples, incubate for 1 hour at 25°C. After that, add two volumes of −20°C acetone and incubate for 20 min at −20°C to remove the biotin-HPDP. At last, Add 15 µl of packed streptavidin-agarose per mg of protein used in the initial protein sample, to purify biotinylated proteins. Incubate the biotinylated proteins with the resin for 1 hour at room temperature. Wash the beads five times with 10 volumes of Neutralization Buffer + NaCl. Centrifuge at 200g for 5 s at room temperature between each wash. Incubate the beads with Elution Buffer to recover the bound proteins. To test for the protein of interest with specific antibodies in SDS-PAGE.

References:

[1].Samie R. Jaffrey and Solomon H. Snyder. The Biotin Switch Method for the Detection of S-Nitrosylated Proteins. Science’s stke.2015.

Biotin-HPDP Dilution Calculator

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Chemical Properties

Cas No. 129179-83-5 SDF Download SDF
Synonyms BiotinHPDP
Chemical Name 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]-N-[6-[3-(pyridin-2-yldisulfanyl)propanoylamino]hexyl]pentanamide
Canonical SMILES C1C2C(C(S1)CCCCC(=O)NCCCCCCNC(=O)CCSSC3=CC=CC=N3)NC(=O)N2
Formula C124H37N5O3S3 M.Wt 539.78
Solubility ≥101.4mg/ml in DMSO Storage Store at -20°C
Shipping Condition Evaluation sample solution : ship with blue ice.All other available size: ship with RT , or blue ice upon request
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.

Features

• Protein labeling—biotinylate antibodies or other proteins for use in protein methods
• Thiol-reactive—reacts with sulfhydryls (-SH), such as the side-chain of cysteine (C)
• Pyridyldithiol-activated—perform reactions at pH 6.5 to 7.5 in buffers such as PBS
• Reversible—forms disulfide bonds, which can be cleaved using DTT or other reducing agent
• Solubility—water insoluble, must be dissolved in DMSO or DMF before further dilution in aqueous buffers
• Medium length—spacer arm is 29.2 angstroms

Background

Biotin-HPDP (N-[6-(biotinamido)hexyl]-3’-(2’-pyridyldithio)propionamide), a sulfhydryl-reactive biotinylation agent, is  a water-insoluble reagent that requires the dissolution of suitable solvents, including dimethyl sulfoxide (DMSO) and dimethylformamide (DMF),  prior to the addition into aqueous reactions. Biotin-HPDP consists of a bicyclic biotin rings structure, a 1,6-diaminohexane spacer group attached to the valeric acid side chain of biotin and a sulfhydryl-reactive group at the end of the spacer. The pyridyl disulfide group at the end of biotin-HPDP is able to react with free thiol groups on proteins and other molecules forming a disulfide bond and releasing pyridine-2-thione. The long spacer arm of biotin-HPDP enables the modified molecule to better bind to the avidin or streptavidin probes.

Reference

Bioconjugate Techniques , 2nd ed. By Greg T.Hermanson  (Pierce Biotechnology, Thermo Fisher Scientific, Rockford, IL).  Academic Press  (an imprint of Elsevier):  London, Amsterdam, Burlington, San Diego . 2008. ISBN 978-0-12-370501-3.