TEV-cut site antibody - 265 003CpH

TEV is a cysteine protease frequently used in biochemistry

pH-sensitive probe fluorescent only under acidic conditions

Rabbit polyclonal purified antibody

Cat. No.: 265 003CpH

Amount: 50 µg

Price: $480.00

Cat. No. 265 003CpH	50 µg specific antibody, lyophilized. Affinity purified with the immunogen, fluorescence-labeled with CypHer5E. CypHer5E^® is a pH-sensitive dye, fluorescent only after antibody internalization at the acidic pH of the lumen of the synaptic vesicle. CypHer5E^® can be detected with the filter sets for Cy5^®. CypHer5E^®: λ_ex 647 nm / λ_em 667 nm This product or portions thereof is manufactured under license from Carnegie Mellon University under U.S. Patent Application Number US 2006-0051758 and related patents. Cy, CyDye, and CypHer are trademarks of GE-Healthcare Ltd. Purchase of reagents related to the CypHer technology from Synaptic Systems GmbH provides a license for non-profit and in-house research use only. Any application of above mentioned technology for commercial purpose requires a separate license from GE-Healthcare Bio-Science Corp., 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA. Albumin was added for stabilization. For reconstitution add 50 µl H₂O to get a 1mg/ml solution in PBS. Either add 1:1 (v/v) glycerol, then aliquot and store at -20°C until use, or store aliquots at -80°C without additives. Reconstitute immediately upon receipt! Avoid bright light when working with the antibody to minimize photo bleeching of the fluorescent dye.
Applications	WB: N/A IP: N/A ICC: 1 : 500 gallery IHC: not tested yet IHC-P: not tested yet Western blot (WB); separation of proteins by PAGE and subsequent transfer to a membrane. Detection of target molecules is carried out with antibodies. Some antibodies require special sample preparation steps. For details, please refer to the “Remarks” section. Immunoprecipitation (IP); Immunoisolation or pulldown of a target molecule using an antibody. For details and product specific hints, please refer to the ”Remarks” section. Immunocytochemistry (ICC) on 4% PFA fixed cells. Immunoreactivity is usually revealed by fluorescence. Some antibodies require special fixation methods. For details, please refer to the “Remarks” section. Immunohistochemistry (IHC) on 4% PFA perfusion fixed tissue with 24h PFA post fixation. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Some antibodies require special fixation methods or antigen retrieval steps. For details, please refer to the ”Remarks” section. Immunohistochemistry (IHC-P) of formalin fixed, paraffin embedded (FFPE) tissue (some antibodies require special antigen retrieval steps, please refer to the ”Remarks” section). Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate.
Label	CypHer5E
Immunogen	Synthetic peptide corresponding to AA 1 to 6 from TEV-cut site
Specificity	TEV-cut protease site is recognized with very strong preference.
Remarks	Excess of extracellular membrane bound SynaptoPhluorin is often removed by TEV protease digestion. This antibody recognizes the neoepitopes generated by TEV cleavage and can be used for membrane internalization studies. The pH sensitive dye regaines its fluorescence after the reacidification of the synaptic vesicle lumen.
Data sheet	265_003cph.pdf

Cat. No.: 265 003CpH

Amount: 50 µg

Price: $480.00

ICC

Background

TEV is a highly specific cysteine protease derived from Tobacco Etch Virus (TEV). The optimum recognition site for this enzyme is the sequence (ENLYFQ(G/S)) and cleavage occurs between the Q and G/S residues leaving a free carboxy-terminus.

1 ) Sun C et al., Protein Expr. Purif. (2012)
2 ) Lundbäck AK et al., Anal. Biochem. (2008)
3 ) Phan J et al., J. Biol. Chem. (2002)
4 ) Kapust RB et al., Biochem. Biophys. Res. Commun. (2002)
5 ) Kapust RB et al., Protein Eng. (2001)

Protocols

ICC

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