Human RAF proto-oncogene serine/threonine-protein kinase (RAF1) ELISA Kit

RAF proto-oncogene serine/threonine-protein kinase (RAF1) is the cellular homolog of viral raf gene (v-raf) . The encoded protein is a MAP kinase kinase kinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated, the cellular RAF1 protein can phosphorylate to activate the dual specificity protein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in RAF1 are associated with Noonan syndrome 5 and LEOPARD syndrome 2.

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Human RAF proto-oncogene serine/threonine-protein kinase (RAF1) ELISA Kit has high sensitivity and excellent specificity for detection of Human RAF1. No significant cross-reactivity or interference between Human RAF1 and analogues was observed.

This Human RAF proto-oncogene serine/threonine-protein kinase (RAF1) ELISA Kit employs a two-site sandwich ELISA to quantitate RAF1 in samples. An antibody specific for RAF1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyRAF1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for RAF1 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of RAF1 bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.