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ABC Transporter Products

GenoMembrane has excellent expertise and experience in the production of various drug transporters.

We provide “ABC Transporter Membranes and Vesicles”, which are prepared from insect Sf9 cells expressing various ABC transporters.

We also provide ready-to-use kit containing all reagents needed for ATPase assay using ABC Transporter Membranes and vesicular transport assay using ABC Transporter Vesicles.

ATPase assay and vesiculare transport assay using our products are helpful to clarify the interactions between a compound of interest and ABC transporters.

ABC Transporter Membranes and Reagent Kit for ATPase Assay

ABC Transporter Membranes

One vial of the membrane producuts can perform 125 ATPase assays.
The protocol and product size (2.5mg／0.5ml／vial) of all the following products is same.

○　Human MDR1 Membranes (GM cat. No. GM0015)

○　Mouse Mdr1a Membranes (GM cat. No. GM0004)

○　Mouse Mdr1b Membranes (GM cat. No. GM0016)

○　Human MRP2 Membranes (GM cat. No. GM0001)

○　Rat Mrp2 Membranes 　　　　 (GM cat. No. GM0002)

○　Dog Mrp2 Membranes 　　　　　 (GM cat. No. GM0014)

○　Monkey Mrp2 Membran (GM cat. No. GM0018)

○　Control Membranes (GM cat. No. GM0003)

Click ATPase assay protocol (ver1.9) for the protocol

* ABC transporter membranes may contain a trace amount of endogenous sensitive ATPase originating from the Sf9 cells. It is, therefore, recommended to conduct an ATPase assay with Control Membranes (Cat. No. GM0003) in addition to the membranes containing the transporter of interest.

Reagent Kit for ABC Transporter Membranes

GenoMembrane provides “Ready-to-use” ATPase Assay kits.

This kit can perform 160 ATPase assays and 16 phosphate standard (one kit full 96-well plate×2).

ABC Transporter ATPase Assay Reagents Kit (GM3020)

[Contents]　・Reaction Buffer

・100 mM Orthovanadate Solution

・200 mM MgATP Solution

・10 mM Phosphate Standard Solution

・100 mM Orthovanadate Solution

・Reductant

・Stop Solution

・Coloring A Solution

・Coloring B Solution

Click (GM3020, ver2.0) for the protocol of reagent kit above

ABC Transporter Vesicles and Reagent Kit for Vesicular Transport Assay

ABC Transporter Vesicles

One vial of the membrane producuts can perform 50 vesicular transport assays.
The product size of all the products below is same. (2.5mg／0.5ml／vial)

○　Human BSEP Vesicles (GM cat. No. GM0005V)

○　Rat Bsep Vesicles (GM cat. No. GM0006V)

○　Dog Bsep Vesicles (GM cat. No. GM0019V)

Click (BSEP, ver2.0) for the protocol of BSEP above

○　Human MRP2 Vesicles (GM cat. No. GM0001V)

○　Rat Mrp2 Vesicles (GM cat. No. GM0002V)

○　Dog Mrp2 Vesicles (GM cat. No. GM0014V)

○　Monkey Mrp2 Vesicles (GM cat. No. GM0018V)

Click (MRP2, ver2.0) for the protocol of MRP2 above

○　Human MRP4 Vesicles (GM cat. No. GM0012V)

○　Rat Mrp4 Vesicles (GM cat. No. GM0020V)

○　Human MRP8 Vesicles (GM cat. No. GM0013V)

○　Human BCRP Vesicles (GM cat. No. GM0008V)

○　Rat Bcrp Vesicles (GM cat. No. GM0007V)

Click (MRP4, MRP8, BCRP protocol, ver2.0) for the protocol of MRP4, MRP8 and BCRP above

○　Control Vesicles (GM cat. No. GM0003 V)

*Because of basal transport activity of the Sf9 cell membrane and nonspecific adsorption of the labeled and non-labeled compound may be observed, depending upon the substrate used. Therefore, we recommend the use of Control Vesicles (Cat.No. GM0003V) as a negative control.

Reagent Kit for ABC Transporter Vesicles New!

GenoMembrane provides two kinds of ready-to-use kit containing all reagents needed for vesicular transport assay using ABC Transporter Vesicles. This kit can perform 100 vesicular transport assays, which is sufficient for two GM vesicles products.

Vesicular Transport Assay Reagents Set

Reagent Set Cat. No. Protocol Contens

For BSEP GM3001 GM3001(ver.1_1)
・Reaction Buffer

・Stopping and Washing Buffer

・10 mM MgATP Solution

・10 mM MgAMP Solution

For MRP2, MRP4, MRP8 and BCRP GM3010 GM3010(ver.1_1)
・Reaction Buffer

・Stopping and Washing Buffer

・10 mM MgATP Solution

・10 mM MgAMP Solution

・200 mM Glutathione Solution

ABC Transporter and Assay

ABC Transporters

The ATP-Binding Cassette (ABC) transporters are a family of membrane proteins, members of which are found in various living organisms from microbes to humans. These transporters have ATPase activity and carry many drugs, fats, sterols and other endogenous metabolites. Forty-seven kinds of ABC transporter have already been identified in the human genome, and approximately 11 kinds are recognized as important drug transporters that transfer materials from the inside of cells to the outside.
ABC transporters may influence the absorption, distribution, metabolism, and excretion (ADME) properties, as well as the efficacy and toxicity of candidate drugs. It is, therefore, important to examine the interaction of candidate drugs with these transporters to help identify drugs with high efficacy and low adverse effects.

ATPase Assay

ABC transporter activity is driven by the energy derived from hydrolysis of ATP.
The ATPase activity increases when a substrate interacts with an ABC transporter. The activity can be estimated by determining the concentration of inorganic phosphate resulting from ATP hydrolysis (ATPase assay). It is possible, therefore, to evaluate the interaction of a drug with an ABC transporter by means of ATPase assay without the need for radioisotope labeling of substrates.
Thus, ATPase assay can be employed as a convenient in vitro screening tool for ABC transporter substrates.

Membrane Vesicle Assay

ABC Transporter Vesicles, which have an inside-out vesicle structure, are prepared from membranes of insect Sf9 cells expressing ABC transporters for membrane vesicle assay. The transporters in these inside-out vesicles import the substrate from outside of the cell membrane vesicles to inside. Subsequently, quantitative measurement with HPLC or LC/MS/MS, or with radioactivity or fluorescence measurement in the case of appropriately labeled compounds, can be used to evaluate directly the transport activity by determining the amounts of the compounds in the cell membrane vesicles.