Technical Data

M2865-03
Clone Type
Polyclonal
Host
Rabbit
Source
Human
Isotype
IgG
Grade
Affinity Purified
Applications
E IP WB
Crossreactivity
Hu Mo Rt
Shipping Temp
Blue Ice
Storage Temp
-20°C
Notes
Preservative Free
Rabbit Anti-MEK1, MEK2, phosphorylated (Ser217/221) (MAPK/ERK Kinase 1, MEK 1, Dual Specificity Mitogen-activated Protein Kinase Kinase 1, MAP Kinase Kinase 1, MAP2K1, MAPKK 1, MKK1, PRKMK1, ERK Activator Kinase 1 / MAPK/ERK Kinase 2, Dual Specificity Mitogen-activated Protein Kinase Kinase 2, MAP Kinase Kinase 2, MAP2K2, MAPKK 2, MKK2, PRKMK2, ERK Activator Kinase 2)
EC=2.7.12.2, Protein Kinase Mitogen Activated Kinase 1, Protein Kinase Mitogen Activated Kinase 2

MEK1 and MEK2, also called MAP or Erk kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation. Activation of MEK1 and MEK2 occurs through phosphorylation of two serine residues at positions 217 and 221 (in the activation loop of subdomain VIII), by Raf-like molecules. MEK1/2 is activated by a wide variety of growth factors and cytokines, and also by membrane depolarization and calcium influx. Constitutively active forms of MEK1/2 are sufficient for the transformation of NIH/3T3 cells or the differentiation of PC12 cells. MEK activates p44 and p42 MAP kinase by phosphorylating both threonine and tyrosine residues at sites located within the activation loop of kinase subdomain VIII. Phospho-MEK1/2 (Ser217/221) selectively recognizes active MEK, i.e., only when phosphorylated at Ser217/221, and hence is an excellent marker of MEK1/2 activity.

Applications
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilution
Western Blot: 1:1000. Incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight. Immunoprecipitation: 1:50 Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunogen
Synthetic phosphopeptide corresponding to residues around Ser217/221 of human MEK1/2 (KLH coupled). MW: 45kD.
Form
Supplied as liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.
Purity
Purified by Protein A affinity chromatography.
Specificity
Detects endogenous levels of MEK1/2 only when activated by phosphorylation at Ser217/221. Does not crossreact with related kinases including activated SEK (MKK4), MKK3 or MKK6. It will also react with MEK1/2 singly phosphorylated at Ser217, but not with MEK1/2 singly phosphorylated at Ser221. Species Crossreactivity: Human, mouse and rat.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. Crews, C.M., et al., Science, 258, 478-480 (1992). 2. Alessi, D.R., et al., EMBO. J., 13, 1610-1619 (1994). 3. Rosen, L.B., et al., Neuron 12, 1207-1221 (1994). 4. Crowley, S., et al., Cell 77, 841-852 (1994). 5. Chow, S., et al., Cytometry, 46 (2), 72-78 (2001). 6. MacNicol, M.C., et al., J. Biol. Chem., 275, 3803-3809 (2000). 7. Pizon, V., et al., Oncogene, 19, 6074-6081 (2000). 8. Rosenberger, C.M., et al., J. Biol. Chem., 277, 18753-18762 (2002). 9. Wang, X., et al., J. Biol. Chem., 275, 39435-39443 (2000).
USBio References
No references available
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