Cell Analysis

Cell Analysis
Cell analysis techniques improve our basic understanding of cellular mechanisms and help identify the proteins and pathways involved in disease.
Millipore provides one of the broadestofferings of multiwall filter plates, antibodies, and other reagent for studying cells. These products and their application are described in detail in a separate publication, the “ImmunodetectionApplication and Product Guide”, which is available upon request from Millipore.
In the chapter that follows, you will find an overview of our products for immunocytochemistry/immunohistochemistry,western blotting, and immunoprecipitation. You will also find protocols for the detection of BrdU and free phosphates, as well as protocols for ELISA assays and cell staining. These procedures representonly a handful of the many techniques that we support. Additional information can be found in the Immunodetection handbook and on our web site.

General Protocol for the Sandwich ELISA Method
1.Before the assay, both antibody preparations should be purified and one must be labeled.
2. For most applications, a polystyrene micro titer plate is best; however, consult manufacturer guidelinesto determine the most appropriate type of plate for protein binding.
3. Bind the unlabeled antibody to the bottom of each well by adding approximately 50 µL of antibody solution to each well (20µg /mL in PBS). PVC will bind approximately 100 ng /well (300 ng/cm2). The amount of antibody used will depend on the individual assay, but if maximal binding is required, use at least 1µg/well. Thisis well above the capacity of the well, but the binding will occur more rapidly, and the binding solution can be saved and used again.
4. Incubate the plate overnight at 4°C to allow completebinding.
5. Wash the wells twice with PBS. A 500mL squirt bottle is convenient. The antibody solution washes can be removed by flicking the plate over a suitable container.
6. The remaining...