Description
The O24-836 monoclonal antibody specifically binds to the human MEK1 pS218 and MEK2 pS222 phosphorylated sites. MEK1 and MEK2 (MEK1/MEK2) are ~45 kDa dual-specificity protein kinases that are also known as MAPK/ERK Kinase 1 and 2 and MAP kinase kinases (aka, MAP2K1 and 2, MAPKK1 and 2, or MKK1 and 2). MEK1 and MEK2 activation is dependent upon phosphorylation of certain amino acid residues including serine 218 (pS218) of MEK1 and serine 222 (pS222) of MEK2 by MAP kinase kinase kinases (MAP3Ks), such as by Raf kinases. MEK1/MEK2 can phosphorylate and thus activate the MAP (Mitogen-Activated Protein) kinases, ERK1 and ERK2. The ERK kinases are critically involved in multiple signal transduction pathways induced by growth factors, cytokines, and mitogens. These kinases regulate cellular growth, differentiation, survival, motility and angiogenesis. Dysregulated activation of MEK1/2 is observed in tumor cells. During antibody development, the purified O24-836 monoclonal antibody was found to detect phosphorylated MEK1/MEK2 by Western blot analysis of cellular lysates and by indirect immunofluorescent staining and flow cytometric analysis of fixed and permeabilized cells. This antibody crossreacts with phosphorylated MEK1/MEK2 expressed by mouse cells as tested by Western blot analysis and immunofluorescent staining and flow cytometric analysis.
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Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. An isotype control should be used at the same concentration as the antibody of interest.
3. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
6. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
7. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
9. Source of all serum proteins is from USDA inspected abattoirs located in the United States.