Biotin-HPDP
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
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.
2. 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
3. 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
4. Sambeeta Das. "DESIGNS FOR DIRECTING MOTION AT THE MICRO-AND NANOSCALE." The Pennsylvania State University.August 2016.
5. Isamar Ortiz‐Rivera, Taylor M.Courtney,Ayusman Sen. "Enzyme Micropump‐Based Inhibitor Assays." Advanced Functional Materials.April 5, 2016.
• 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
| Physical Appearance | A solid |
| Storage | Store at -20°C |
| M.Wt | 539.78 |
| Cas No. | 129179-83-5 |
| Formula | C124H37N5O3S3 |
| Synonyms | BiotinHPDP |
| Solubility | ≥101.4mg/mL in DMSO, ≥8.29 mg/mL in EtOH with ultrasonic, <2.51 mg/mL in H2O |
| 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 |
| SDF | Download SDF |
| Canonical SMILES | C1C2C(C(S1)CCCCC(=O)NCCCCCCNC(=O)CCSSC3=CC=CC=N3)NC(=O)N2 |
| 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. |
| Biotinylation method [1]: | |
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Sample |
S-nitrosylated proteins |
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Preparation method |
Soluble in DMSO or DMF. |
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Reaction Conditions |
50mM, 25 ℃ for 1 h |
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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. |
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References: [1].Samie R. Jaffrey and Solomon H. Snyder. The Biotin Switch Method for the Detection of S-Nitrosylated Proteins. Science’s stke.2015. |
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| Description | Biotin-HPDP is a sulfhydryl-reactive biotinylation agent. | |||||
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Quality Control & MSDS
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