Biotin-HPDP |
| Catalog No.GC11037 |
Biotin-HPDP is a sulfhydryl-reactive biotinylation reagent that forms a reversible disulfide linkage.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 129179-83-5
Sample solution is provided at 25 µL, 10mM.
Biotin-HPDP is a sulfhydryl-reactive biotinylation reagent that forms a reversible disulfide linkage.Biotin-HPDP consists of a bicyclic Biotin ring structure, a 1, 6-diaminohexane attached to the side chain of biotin valonate, and a sulfhydryl reactive group at the end of the side chain[1]. It is used to label protein cysteines and other substrates that contain sulfhydryl groups.
After NEM alkylation, certain (but not all) residual free cysteine residues may react with biotin-HPDP, resulting in the coisolation of proteins containing such cysteine residues[2].
A kinetic study of the modification reactions that generate monothiophosphate disulfide linkages with either 5'-GMPS alone or 5'-GMPS-primed RNA as the substrate revealed that the second-order rate constants increased as the pH was decreased. For example, when the reaction pH was lowered from 8 to 4, the k2 value for the coupling reaction between N-(6-[biotinamido]hexyl)-3'-(2'-pyridyldithio)propionamide (biotin-HPDP) and GMPS increased 67-fold from 1.84 to 123 M(-1) x s(-1) [3].
References:
[1]: 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]: Zhou B, Wang Y, Yan Y, Mariscal J, Di Vizio D, Freeman MR, Yang W. Low-Background Acyl-Biotinyl Exchange Largely Eliminates the Coisolation of Non-S-Acylated Proteins and Enables Deep S-Acylproteomic Analysis. Anal Chem. 2019 Aug 6;91(15):9858-9866. doi: 10.1021/acs.analchem.9b01520. Epub 2019 Jul 11. PMID: 31251020; PMCID: PMC7451198.
[3]: Wu CW, Eder PS, Gopalan V, Behrman EJ. Kinetics of coupling reactions that generate monothiophosphate disulfides: implications for modification of RNAs. Bioconjug Chem. 2001 Nov-Dec;12(6):842-4. doi: 10.1021/bc0100612. PMID: 11716671.
| 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. |
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| Cas No. | 129179-83-5 | SDF | |
| 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 | C24H37N5O3S3 | M.Wt | 539.78 |
| Solubility | 3mg/mL in DMSO | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.8526 mL | 9.263 mL | 18.5261 mL |
| 5 mM | 0.3705 mL | 1.8526 mL | 3.7052 mL |
| 10 mM | 0.1853 mL | 0.9263 mL | 1.8526 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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μL
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Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
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