MembranePro™ Functional Protein Expression (FPE) System

For researchers studying G protein–coupled receptors (GPCRs) and other membrane proteins, the MembranePro™ Functional Protein Expression (FPE) System delivers enriched, functional membrane proteins efficiently and reliably. Key features of the system:

  • Proteins are displayed on human (or other mammalian) cell membranes
  • Cellular quality control and mammalian posttranslational processing helps produce functional protein
  • Proteins bud off as lipoparticles and are easily collected from the culture medium
  • Lipoparticles are enriched for expressed receptors

Improved Workflow

The MembranePro™ FPE System workflow is less labor-intensive than traditional production of membrane fractions. Cloning and transfection have been optimized via TOPO® cloning vectors and Lipofectamine® 2000 Transfection Reagent, respectively, and there is no need for laborious ultracentrifugation steps. Briefly, the MembranePro™ protocol requires the following:

  1. Clone your gene of interest into a recommended TOPO® vector for mammalian expression, transform, and prepare plasmid.
  2. Transfect 293FT cells with your TOPO® cloned vector and MembranePro™ Reagent.
  3. After a 48 hr incubation, the lipoparticles are harvested from the culture supernatant and precipitated with the MembranePro™ Precipitation Mix.
  4. Store lipoparticles at –80°C or proceed to downstream biochemical assays, including receptor–ligand binding studies.

Soluble, Functional Proteins Ready for Use

The MembranePro™ FPE System provides an easy method to harvest soluble, functional membrane proteins that are easily adaptable to your current downstream assays. Additional benefits include:

  • Harvested lipoparticles contain overexpressed membrane proteins in a homogeneous preparation (easy to pipet, not clumpy)
  • No need to establish stable cell lines expressing your proteins of interest
  • Eliminate concerns about down-regulation of expression due to toxicity
  • Avoid capital investment and maintenance of an ultracentrifuge

Product Configurations

MembranePro™ products are offered in two configurations—as an Expression Kit (includes pEF6/V5 His-TOPO® vector, MembranePro™ Reagent, 293FT cells, Lipofectamine® 2000 Transfection Reagent, MembranePro™ Precipitation Mix) or as a Support Kit (includes Lipofectamine® 2000 Transfection Reagent, MembranePro™ Reagent, MembranePro™ Precipitation Mix). The Expression Kit enables 10 reactions (sufficient reagents for 10 transfections of 293FT cells in T-175 flasks), and the Support Kit is available in 10, 60, and 600 reaction sizes.
The resulting lipoparticles from a single reaction generally contain enough material to produce up to 1,000 data points in ligand-binding assays (e.g., ligand-binding assays in 10 x 96-well plates). The number of data points will vary depending on the expression level of the membrane protein being studied.