Catalog # PK6930


Profilin Phospho-Regulation

Antibody Sampler Kit

$395

Profilins are small actin-binding proteins that have functions in cell motility, cytokinesis, gene transcription, endocytosis and neuronal plasticity. Four profilin isoforms have been identified in mammals. Profilin-1 (PFN1) and profilin-2a (PFN2a) isoforms are highly conserved in structure, but PFN1 is ubiquitously expressed while PFN2a is preferentially enriched in brain. In addition, there are two testis-specific profilins, PFN3 and PFN4, that significantly differ in primary sequence and function compared to PFN1 and PFN2a. Profilin is phosphorylated at both tyrosine and serine residues in vivo. Tyr-129 is phosphorylated in response to VEGF-A stimulation, and this promotes profilin actin binding and polymerization. Tyr-129 phosphorylation may be important for angiogenesis induced by injuries. Ser-138 is phosphorylated by ROCK and dephosphorylated by PP1. This serine phosphorylation inhibits G-actin binding, as well as decreases profilin's aggregation suppressor activity by inhibiting binding to huntingtin. Thus, Tyr-129 phosphorylation may activate while Ser-138 phosphorylation may inhibit profilin activity.

References
Shao, J. et al. (2008) Mol Cell Biol. 28(17):5196.
Fan, Y. et al. (2012) Nat Cell Biol. 14(10):1046.
Shao, J. & Diamond, M.I. (2012) PLoS ONE 7(3): e32802.
Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.



*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

PP4821 Stanslowsky N. et al. (2016) J Neurosci. 36(47):12027. (WB: human striatal medium spiny neurons)
PP4791 Stanslowsky N. et al. (2016) J Neurosci. 36(47):12027. (WB: human striatal medium spiny neurons)
PP4751 Dombert B. et al. (2017) Front Mol Neurosci. 10:346. (ICC: mouse spinal motoneurons)
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. (WB, ICC: mouse intestinal myofibroblasts)
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. (Western Blot)

Western blot of HUVEC stimulated with Pervanadate (1 mM) for 30 min (lanes 1-4). The blots were treated with alkaline phosphatase to remove phosphorylation (lanes 2 & 4), then probed with anti-Profilin (a.a. 126-137) (lanes 1 & 2) or anti-Profilin (Tyr-129) phospho-specific (lanes 3 & 4). The antibodies detect profilin at 15 kDa. In lanes 3 & 4, the antibody also detects an unknown 20 kDa protein.

Western blot of Jurkat stimulated with calyculin A (100 nM) for 30 min (lanes 1-6). The blots were probed with anti-Profilin (a.a. 132-140) (lanes 1-3) or anti-Profilin (Ser-138) phospho-specific (lanes 4-6). Both antibodies were used in the absence (lanes 1 & 4) or presence of unphosphorylated Profilin (Ser-138) (lanes 2 & 5) or phospho-Profilin (Ser-138) (lanes 3 & 6) blocking peptides.



This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
Profilin (a.a. 132-140) Rabbit pAb50 μlWB, EHu, Rt, Ms15
Profilin (Ser-138), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms15
Profilin (a.a. 126-137) Rabbit pAb 50 μlWB, EHu, Rt, Ms15
Profilin (Tyr-129), phospho-specific Rabbit pAb 50 μlWB, EHu, Rt, Ms15
Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The Profilin phospho-regulation antibody sampler kit can be used to detect phosphorylation of Tyr-129 and Ser-138 relative to total profilin expression levels. The kit includes rabbit polyclonal phospho-specific antibodies to Tyr-129 and Ser-138, as well as rabbit polyclonal antibodies that detect different epitopes in the C-terminal region of Profilin. The kit also includes an anti-Rabbit Light Chain specific:HRP secondary reagent for detection of rabbit polyclonal antibody in western blot, ELISA, or immunocytochemistry.