Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Phospho-MARK Family (Activation Loop) Antibody #4836

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Endogenous 80 to 95 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-MARK Family (Activation Loop) Antibody detects endogenous levels of phosphorylated MARK family members, MARK1 at threonine 215, MARK2 at threonine 208, and MARK3 at threonine 234 (A.K.A. 211 in isoforms 3-6) . This antibody does not react with MARK4.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding threonine 215 of human MARK1. Antibodies were purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells transfected with either wild-type or threonine to alanine mutations at the respective phosphorylation sites of the MARK family members, using Phospho-MARK Family (Activation Loop) Antibody. Transfected constructs contain a GST and HA tag. Transfection efficiency monitored with HA Antibody, #2367.

Western Blotting

Western Blotting

Western blot analysis of extracts from Raji and BaF3 cells, showing endogenous levels of phosphorylated MARK family members, using Phospho-MARK Family (Activation Loop) Antibody.

Background

Microtubule associated proteins regulate the stability of microtubules and control processes such as cell polarity/differentiation, neurite outgrowth, cell division and organelle trafficking (1). The MARK (MAP/microtubule affinity-regulating kinases) family (MARK1-4) of serine/threonine kinases was identified based on their ability to phosphorylate microtubule-associated proteins (MAPs) including tau, MAP2 and MAP4 (2-6). MARK proteins phosphorylate MAPs within their microtubule binding domains, causing dissociation of MAPs from microtubules and increased microtubule dynamics (2-4). In the case of tau, phosphorylation has been hypothesized to contribute to the formation of neurofibrillary tangles observed in Alzheimer's disease. Overexpression of MARK leads to hyperphosphorylation of MAPs, morphological changes and cell death (4). The tumor suppressor kinase LKB1 phosphorylates MARK and the closely related AMP-kinases within their T-loops, leading to increased activity (7).

  1. Drubin, D.G. and Nelson, W.J. (1996) Cell 84, 335-44.
  2. Illenberger, S. et al. (1996) J Biol Chem 271, 10834-43.
  3. Drewes, G. et al. (1995) J Biol Chem 270, 7679-88.
  4. Drewes, G. et al. (1997) Cell 89, 297-308.
  5. Kato, T. et al. (2001) Neoplasia 3, 4-9.
  6. Trinczek, B. et al. (2004) J Biol Chem 279, 5915-23.
  7. Lizcano, J.M. et al. (2004) EMBO J 23, 833-43.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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