Product Pathways - MAPK Signaling
Phospho-MEK1/2 (Ser221) Blocking Peptide #1210
Description
This peptide is used to specifically block Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb #2338 reactivity.
Quality Control
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb #2338 signal in both immunohistochemistry and Western blotting.
Western Blotting
Western Blot analysis of extracts from NIH/3T3 cells, untreated or serum-treated, using Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb #2338 (left) or same antibody preincubated with Phospho-MEK1/2 (Ser221) Blocking Peptide (right)
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb #2338 in the presence of control peptide (left) or Phospho-MEK1/2 (Ser221) Blocking Peptide (right).
Applications
Use as a blocking reagent to evaluate the specificity of antibody reactivity in Western immunoblotting and immunohistochemistry protocols.
Directions for Use
For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 ul total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb #2338 data sheet. For Western blotting, add 10 ul of antibody and 10 ul of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 30 minutes before allowing to react with the blot.
Background
MEK1 and MEK2, also called MAPK or Erk kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation (1-3). Activation of MEK1 and MEK2 occurs through phosphorylation of two serine residues at positions 217 and 221, located 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 (1-4). Constitutively active forms of MEK1/2 are sufficient for the transformation of NIH/3T3 cells or the differentiation of PC-12 cells (4). 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.
- Crews, C.M. et al. (1992) Science 258, 478-480.
- Alessi, D.R. et al. (1994) EMBO J. 13, 1610-1619.
- Rosen, L.B. et al. (1994) Neuron 12, 1207-1221.
- Cowley, S. et al. (1994) Cell 77, 841-852.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
For Research Use Only. Not For Use In Diagnostic Procedures.
