In Cell ELISA Kit

A fast, robust, and efficient method for quantifying protein levels directly in cells

The MaxSuppressor™ In Cell ELISA Kit is an enzyme immunoassay for quantitative protein analysis directly in cell cultures without the need for cell lysis or protein gels. The technique can be thought of as an ELISA performed on fixed, permiabilized cells rather than on an immobilized antigen.  As in traditional ELISA assays, this kit detects protein using a series of two antibodies, a primary antibody for the protein of interest and an enzyme-conjugated secondary antibody.  This kit includes mouse primary antibody targeting glyceraldehyde 3-phosphate dehydrogenase (GAPDH) or the researcher can use their own primary antibody.  Both anti-mouse and anti-rabbit HRP conjugated secondary antibodies are included in the kit so that primary antibodies from either mouse or rabbit may be used for detection of the protein of interest.  The protein antibody complex is visualized by adding 3,3'5,5 ۥ-Tetramethylbenzidine (TMB), a substrate for the HRP, that turns blue when oxidized with hydrogen peroxide and is subsequently changed to yellow by adding sulfuric or phosphoric acid. Finally, the assay results are collected in a spectrophotometric plate reader at 450 nm. The relative amount of protein in the cells will be directly proportional to the amount of signal that is obtained, making this a simple, convenient, and accurate technique for quantitating protein levels within cells.

Kit Features:

1. Results are obtained in as few as 3 hours instead of the 12 - 24 hours needed for western analysis.
2. Protein is measured directly in cells; no need to extract protein or run gels.
3. Users are able to rapidly quantitate protein levels after treating cultured cells with drugs or small RNA molecules such as miRNA or siRNA.
4. Ideal for cellular screening assays. For example, quantify levels of phosphorylated protein that can reflect the overall state of cell cultures.
5. Allows users to measure GAPDH levels for all of your experimental needs.
6. Contains all of the materials for washing, fixing, and permiabilizing cells as well as all of the other needed reagents such as antibody dilution solutions, concentrated PBS, wash solution, enzyme substrate, and Stop Solution.

Figure Legend. SKOV3 cells (6,000 cells/well), a cervical cancer cell line, were reverse transfected with either a siRNA targeting GAPDH or a negative control siRNA using 0.6 ul of hyperfect (Qiagen) in a 96-well plate. 72 hours after transfection, the MaxSuppressorTM In Cell ELISA Kit was used to detect GAPDH knockdown.

 

Reference	Description	Qté	Prix unitaire Euros HT
3440-02	MaxSuppressor™ In Cell ELISA Kit	1 kit, 192 réactions	357.00 EUR