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Phosphorylation ELISA Kits
Semi-Quantitative, Sandwich ELISAs for Detection of Phosphorylated Proteins in Cell Lysates
RayBio® Human PhosphoELISA (Enzyme-Linked Immunosorbent Assay) kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phosphorylated protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blot analysis.
For a comparison & overview of all our ELISA systems, click here.
How it works
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of phosphorylated protein. An specific capture antibody (anti-pan-antibody or anti-phosphorylated antibody) has been coated onto a 96-well plate. Samples are pipetted into the wells and target protein present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phosphorylated antibody or anti-pan-antibody is used to detect target protein. After washing away unbound antibody, secondary antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of target bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
1. Recombinant Human EGF Stimulation of A431 Cell Lines:
- A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using this phosphoELISA and Western Blot.
- The A431 cells were treated with 100 ng/mL recombinant human EGF for 20 minutes to induce phosphorylation of EGF R. Serial dilutions of lysates were analyzed in this ELISA and by Western blot. Immunoblots were incubated with anti-phospho-EGFR (Tyr 1086).
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