Members of the CRMP family were discovered independently in different species by several groups working in parallel. Among the five members of the family, CRMP-2 was first identified in 1995. Group of researchers led by Goshima found out that CRMP-2 played a role in the transduction of the extracellular Semaphorin 3A (Sema3A), an inhibitory protein for axonal guidance in chick dorsal root ganglion (DRG). The protein was first named as CRMP-62 having a relative molecular mass of 62 kDa and later referred as CRMP-2. Concurrently, a 64 kDa protein named as TOAD-64 for Turned On After Division, was shown to increase significantly during the development of the cortex of the brain. The cDNA sequence of TOAD-64 corresponded to that of rat CRMP-2. In 1996, mouse CRMP-4, often referred to as Ulip for Unc-33 like phosphoprotein, was discovered by Byk and colleagues, using a rabbit polyclonal antiserum which recognized a 64 kDa mouse brain specific phosphoprotein. In the same year, several other studies cloned CRMPs-1-4 in rat and dihydropyrimidinase (DHPase) homologous sequence of CRMPs-1, -2, and -4 in human fetal brain. Finally, in 2000, CRMP-5 was discovered using two-hybrid screenings of brain libraries or purification from a proteic complex. In following researches, CRMPs were studied as target antigens for autoantibodies in various autoimmune neurodegenerative disorders.
Background References
1. Charrier E.; Reibel S.; Rogemond V.; Aguera M.; Thomasset N.; Honnorat J. (August 2003). "Collapsin response mediator proteins (CRMPs) - Involvement in nervous system development and adult neurodegenerative disorders". Molecular Neurobiology. 28 (1): 51–63.
2. Hou ST, Jiang SX, Smith RA (2008). Permissive and repulsive cues and signaling pathways of axonal outgrowth and regeneration. International Review of Cell and Molecular Biology. Vol. 267. pp. 125–181.
Western blot analysis of Phospho-CRMP2 (T514) on different lysates with Rabbit anti-Phospho-CRMP2 (T514) antibody (HA721674) at 1/1,000 dilution.
Lane 1: PC-12 cell lysate (20 µg/Lane) Lane 2: Human brain tissue lysate (30 µg/Lane) Lane 3: PC-12 treated λpp for 1 hour cell lysate (20 µg/Lane)
Predicted band size: 62 kDa Observed band size: 70 kDa
Exposure time: 45 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721674) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Cell treatment (CT)
Western blot analysis of Phospho-CRMP2 (T514) on different lysates with Rabbit anti-Phospho-CRMP2 (T514) antibody (HA721674) at 1/1,000 dilution.
Lane 1: Mouse brain tissue lysate Lane 2: Mouse brain tissue lysate, the membrane treated with λpp for 1 hour
Lysates/proteins at 30 µg/Lane.
Predicted band size: 62 kDa Observed band size: 70 kDa
Exposure time: 5 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721674) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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