EnzyFluo™ Custom Cell-Based ELISA Kit

Protocol SDS

Application

• A reagent kit that allows our customers to develop their own cell-based ELISA. Determination of relative protein content and signal pathway (e.g. phosphorylation status) in whole cells. Evaluation of target modulation by drugs, activators, inhibitors, siRNA etc. All that is required is a primary mouse antibody (Ab1) for the specific target of interest. (BioAssay Systems also offers custom development. Please inquire).

Key Features

• New and improved. Total assay time reduced from the standard 21 hours to 6.5 hours (hands-on time 2.5 hrs).
• Simple and convenient. Cells are directly cultured in 96-well plates. No cell lysis necessary.
• Accurate and high-throughput. Target protein is normalized to total cellular protein in the same well, greatly minimizing well-to-well variations. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Method

• Cell-Based ELISA (FL530/585 nm, 360/450 nm)

Samples

• Cells

Species

• human, mouse, rat and more, depending on choice of primary antibody

Procedure

• Assay takes 6.5 hrs, hands-on time 2.5 hrs

Size

• 100 tests

Shelf Life

• 6 months

More Details

Many biological processes are regulated by changing the concentration and/or activity of proteins in the cell. A rapid assay capable of monitoring such changes is highly desirable in life science research and drug discovery. BioAssay Systems’ Custom Cell-based ELISA Kit provides a rapid and sensitive starter tool for measuring the relative content of any target protein in cultured cells. Only a target-specific, primary mouse antibody (Ab1) for the protein of interest is needed to run the assay. This simple and efficient assay eliminates the need for cell lysate preparation and can be used to determine a particular protein content, or signaling pathway activity and effects of treatments by e.g. drugs, inhibitors, siRNA or activators. In this assay, cells grown in a 96-well plate are fixed and permeabilized. The target protein is measured using your own Ab1 followed by a HRP-conjugated secondary antibody with a fluorogenic substrate, and a fluorescent reagent that measures the total protein in the same well. This convenient kit provides all necessary reagents for the user to develop their own assay. Upon request, BioAssay Systems can also custom develop it for the user. If your primary antibody is raised in rabbit, use our Custom Cell-based ELISA Kit (EFC2R).