pCLIPf-Cox8A Control Plasmid

  • This product was discontinued on 01/01/2013
Catalog #SizeConcentrationPriceQtyAdd to Cart
  
Categories:
Discontinued Products

Description

This control plasmid contains the gene encoding the Cytochrome C oxidase, subunit 8-2 (COX8-2) protein cloned upstream of the CLIPf coding sequence in pCLIPf, as a fusion to the N- terminus of the CLIP-tag. Cytochrome C oxidase is located in the inner mitochondrial membrane and is the terminal enzyme of the respiratory chain. The COX8-2-CLIPf fusion protein gives mitochondrial fluorescence when labeled with CLIP-Cell™ substrates. The full sequence and map for pCLIPf-Cox8A can be downloaded.

The CLIP-tag™ protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. CLIP-tag is a protein tag based on human O6-alkylguanine-DNA-alkyltransferase (hAGT). CLIP-tag substrates are derivatives of benzylcytosine (BC). In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the reactive cysteine of CLIP-tag forming a stable thioether bond. Although CLIP-tag is based on the same protein as SNAP-tag, the benzylcytosine substrates form a separate class of substrates, different from the benzylguanine substrates recognized by SNAP-tag. CLIP-tag and SNAP-tag® can be used for orthogonal and complementary labeling of two proteins simultaneously in the same cells.

pCLIPf-Cox8A contains an improved version of CLIP-tag, termed CLIPf. CLIPf displays faster kinetics in in vitro labeling and fast, specific and efficient labeling in live and fixed cell applications, thereby rendering it a desired research tool for analysis of protein dynamics. 

There are two steps to using this system: sub cloning and expression of the protein of interest as a CLIPf fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expressing the CLIPf-Cox8A fusion protein is described in this document. The labeling of the fusion proteins with CLIP-tag substrates is described in the instructions supplied with CLIP-tag substrates.

Figure 1: Live HEK293 cells transiently transfected with pCLIPf-Cox8A. Cells were labeled with CLIP-Cell TMR-Star (red) for 30 minutes and counterstained with Hoechst 33342 (blue).


N9217b_thumb

Properties and Usage

Materials Required but not Supplied

Cell culture media and reagents
Mammalian cell lines
Transfection reagents
CLIP-tag substrates

Storage Temperature

-20°C

Sequence Files

Fasta  GenBank  

Notes

  1. Storage: pCLIPf-Cox8A is supplied in TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA) at a concentration of 0.5 µg/µl. Plasmid solutions can be stored at 4°C for up to one week. For long-term storage -20°C is recommended.

FAQs

  1. Cellular Imaging and Analysis FAQs

Tech Tips

If you generate more plasmid DNA by bacterial transformation, we recommend isolating the plasmid DNA using an endotoxin-free plasmid prep kit prior to transfection into mammalian cells.

Protocols

  1. Expression of CLIPf-Cox8A (N9217)

Selection Charts

Troubleshooting Guides

Interactive Tools

Application Notes

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.