Revision 4
#41343
Store at -20C
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
PTMScan
Storage
Species predicted to react based on 100% sequence homology
All Species Expected
Description
Biotin, also known as vitamin B7, is a chemical moiety that can be post-tranlationally added to proteins via a process known as biotinylation. Biotinylated proteins and peptides bind very tightly to avidin groups. Biotinylation of protein targets can be achieved using chemical or enzymatic methods.
PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology (CST) for peptide enrichment by immunoprecipitation using a specific bead-conjugated antibody in conjunction with liquid chromatography (LC) tandem mass spectrometry (MS/MS) for quantitative profiling of post-translational modification (PTM) sites in cellular proteins. These include phosphorylation (PhosphoScan®), ubiquitination (UbiScan®), acetylation (AcetylScan®), and methylation (MethylScan®), among others. PTMScan® Technology enables researchers to isolate, identify, and quantitate large numbers of post-translationally modified cellular peptides with a high degree of specificity and sensitivity, providing a global overview of PTMs in cell and tissue samples without preconceived biases about where these modified sites occur (1). For more information on PTMScan® Proteomics Services, please visit www.cellsignal.com/services/index.html
Background
Biotinylation of proteins and peptides has been used for many years and has become a common way to biochemically tag targets of interest (1). One advantage is the very strong binding interaction with avidin that can be used to purify biotin-tagged proteins (2,3). It can also be used as a method for conjugation of a variety of tags such as dyes, beads and other solid substrates for a variety of assays and tools (4,5).
New assays have been developed that improve elucidation of protein subcellular context, localization, and protein-protein interactions. These new cellular assays utilize biotin ligase or engineered ascorbate peroxidase (APEX) fusion proteins that are able to biotinylate nearby protein targets within specific cellular compartments. This type of biotin proximity assay can provide improved information about subcellular localization, multi-subunit components, and recruitment of unknown targets. Using CST's in house developed anti-biotin rabbit monoclonal antibody; we have developed a robust procedure for immunoaffinity enrichment followed by mass spectrometry analysis using CST’s proprietary PTMScan(R) protocols. This allows for identification of biotinylated targets and investigation of new protein interactions. This assay provides rich data for these types of biotin proximity assays such as the APEX (6,7) engineered cell system and other related methods.
Background References
- Alderton, G. et al. (1945) Science 101, 151-2.
- Wright, L.D. et al. (1947) Proc Soc Exp Biol Med 64, 150-3.
- Cronan, J.E. (1990) J Biol Chem 265, 10327-33.
- Hofmann, K. and Kiso, Y. (1976) Proc Natl Acad Sci U S A 73, 3516-8.
- Shamsuddin, A.M. and Harris, C.C. (1983) Arch Pathol Lab Med 107, 514-7.
- Paek, J. et al. (2017) Cell 169, 338-349.e11.
- Hung, V. et al. (2016) Nat Protoc 11, 456-75.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
AcetylScan is a registered trademark of Cell Signaling Technology, Inc.
MethylScan is a registered trademark of Cell Signaling Technology, Inc.
PhosphoScan is a registered trademark of Cell Signaling Technology, Inc.
UbiScan is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 4
#41343
PTMScan® Anti-Biotin Kit
Side by side comparison of two commercially available anti-biotin polyclonal antibodies with Cell Signaling Technology's anti-biotin (A7C2A) rabbit monoclonal antibody. Mouse liver peptides were chemically biotinylated and 1 μg was mixed with 1 mg of untreated mouse liver peptides for each sample. Immunoprecipitated peptides from each of the three antibodies were then analyzed by OrbiTrap™ LC-MS and the average number of biotinylated peptides identified is shown.