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Biotin-HPDP

Catalog No.
A8008
Sulfhydryl-reactive biotinylation reagent, pyridyldithiol-activated
Grouped product items
SizePriceStock Qty
10mM (in 1mL DMSO)
$55.00
In stock
10mg
$50.00
In stock
50mg
$190.00
In stock
100mg
$230.00
In stock
For scientific research use only and should not be used for diagnostic or medical purposes.

Tel: +1-832-696-8203

Email: [email protected]

Worldwide Distributors

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.

Product Citation

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

Chemical Properties

Physical AppearanceA solid
StorageStore at -20°C
M.Wt539.78
Cas No.129179-83-5
FormulaC24H37N5O3S3
SynonymsBiotinHPDP
Solubilityinsoluble in H2O; ≥101.4 mg/mL in DMSO; ≥8.29 mg/mL in EtOH with ultrasonic
Chemical Name5-[(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
SDFDownload SDF
Canonical SMILESC1C2C(C(S1)CCCCC(=O)NCCCCCCNC(=O)CCSSC3=CC=CC=N3)NC(=O)N2
Shipping ConditionSmall Molecules with Blue Ice, Modified Nucleotides with Dry Ice.
General tips We do not recommend long-term storage for the solution, please use it up soon.

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.

Biological Activity

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

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