Cell Signaling Technology

Product Pathways - Neuroscience

p35/25 (C64B10) Rabbit mAb #2680

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IF-F H M R Endogenous 25, 35 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-F=Immunofluorescence (Frozen)
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

p35/25 (C64B10) Rabbit mAb detects endogenous levels of total p35 protein. The antibody also detects endogenous p25 resulting from calpain-mediated cleavage upon neurotoxic insult.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide from the carboxy terminus of human p35.

Western Blotting

Western Blotting

Western blot analysis of extracts from mouse brain, rat brain and human cerebellum using p35/p25 (C64B10) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from mouse and rat brain and wild-type mouse and p35 knock-out mouse brain using p35/25 (C64B10) Rabbit mAb (upper) and CDK5 Antibody #2506 (lower). Matching wild-type and p35 knock-out mouse brain was kindly provided by Dr. Li-Huei Tsai, Massachusetts Institute of Technology, Cambridge, MA.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin embedded human meningioma using p35/25 (C64B10) Rabbit mAb #2680.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human astrocytoma using p35/25 (C64B10) Rabbit mAb #2680 in the presence of control peptide (left) or antigen specific peptide (right).

IF-F

IF-F

Confocal immunofluorescent analysis of cerebellum from normal (left) or p35 knockout (right) mice using p35/25 (C64B10) Rabbit mAb (green). Blue pseudocolor =DRAQ5® #4084 (fluorescent DNA dye). Matching wild-type and p35 knock-out mouse brain was kindly provided by Dr. Li-Huei Tsai, Massachusetts Institute of Technology, Cambridge, MA.

Background

Cyclin-dependent kinases (CDKs) are serine/threonine kinases that are activated by cyclins and govern eukaryotic cell cycle progression. While CDK5 shares high sequence homology with its family members, it is thought mainly to function in postmitotic neurons, regulating the cytoarchitecture of these cells. Analogous to cyclins, p35 and p39 associate with and activate CDK5 despite the lack of sequence homology. CDK5 is ubiquitously expressed, but high levels of kinase activity are detected primarily in the nervous system due to the narrow expression pattern of p35 and p39 in post-mitotic neurons. A large number of CDK5 substrates have been identified although no discrete substrates have been attributed as a function of p35 vs. p39. Amongst many, substrates of CDK5 include p35 and p39. p35 is rapidly degraded (T1/2 <20 min) by the ubiquitin-proteasome pathway (1). However, p35 stability increases as CDK5 kinase activity decreases, and this is likely a result of decreased phosphorylation of p35 at Thr138 by CDK5 (2). NGF activates Erk and EGR1, and induces p35 expression in PC12 cells (3). Proteolytic cleavage of p35 by calpain produces p25 upon neurotoxic insult, resulting in prolonged activation of CDK5 by p25. Accumulation of p25 is found in neurodegenerative diseases such as Alzheimer's disease and Amyotrophic Lateral Sclerosis (ALS) (4-5).

  1. Dhavan, R. and Tsai, L.H. (2001) Nat. Rev. Mol. Cell Biol. 2, 749-759.
  2. Patrick, G.N. et al. (1998) J. Biol. Chem. 273, 24057-24064.
  3. Harada, T. et al. (2001) Nat. Cell Biol. 3, 453-459.
  4. Lee, M.S. et al. (2000) Nature 405, 360-364.
  5. Kusakawa, G. et al. (2000) J. Biol. Chem. 275, 17166-17172.

Application References

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

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