Catalog # AK7710

Actin Phospho-Regulation Staining Kit

Immunocytochemistry Kit

DATASHEET  




$405

Actin is a major cytoskeletal protein involved in diverse cellular functions including cell motility, adhesion, and morphology. Six different actin isoforms have been identified in vertebrates. There are four α isoforms: skeletal, cardiac, and two smooth muscle (enteric and aortic) actins, along with two cytoplasmic actins (β and γ). Actin exists in two principal forms, globular monomeric (G) actin, and filamentous polymeric (F) actin. The assembly and disassembly of actin filaments, and also their organization into functional networks, is regulated by a variety of actin-binding proteins (ABPs). Phosphorylation may also be important for regulating actin assembly and interaction with ABPs. In Dictyostelium, phosphorylation of Tyr-53 occurs in response to cell stress and this phosphorylation may alter actin polymerization. In B cells, SHP-1 tyrosine dephosphorylation of actin leads to actin filament depolymerization following BCR stimulation.

References
Jungbluth, A. et al. (1995) FEBS Let. 375:87.
Baba, T. et al. (2003) J. Immunol. 170: 3762.
Winder, S.J. et al. (2005) J. Cell Sci. 118:651.
Liu, X. et al. (2006) Proc Nat Acad Sci U S A. 103(37):13694.

Immunocytochemical labeling of phosphorylated actin in paraformaldehyde fixed and acetone permeabilized C2C12 cells treated with pervanadate. The cells were labeled with mouse monoclonal anti-Actin (AM2021) and rabbit polyclonal anti-Actin (AP1651) or anti-Actin (Tyr-53) (AP1671). The antibodies were detected using Goat anti-Mouse DyLight® 594 or Goat anti-Rabbit DyLight® 488.



Mouse monoclonal, rabbit polyclonal, and secondary reagents are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store 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:

AM2021 Dutta, P et al. (2014) J Neurochem. 130(3):360 WB: rat brain
AM2021 Muirhead, G et al. (2014) J Mol Neurosci. 53(1):125 WB: rat brain
AM2021 Pritchard, AJ et al. (2014) PLoS One. 9(6):e99444 WB: mouse splenocytes
AP1671 Vonach, C. et al. (2011) British J Cancer. 105:263. WB: human endothelial cells

This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYICC DILUTION
Actin (N-terminal region) Rabbit pAb50 μlWB, E, IP, ICC, IHCHu, Rt, Ms, Ck1:50
Actin (Tyr-53), phospho-specific Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms, Ck1:50
Actin (C-terminal region) Mouse mAb50 μlWB, E, ICC, IHCHu, Rt, Ms, Ck1:50
Anti-Mouse Ig:DyLight®594 Goat pAb100 μlICC, IHCMs1:200
Anti-Rabbit Ig:DyLight®488 Goat pAb100 μlICC, IHCRb1:200
KIT SUMMARY

The actin phospho-regualtion kit can be used to label actin phosphorylation at Tyr-53 relative to total actin expression pattern. The kit contains anti-Actin (Tyr-53) phospho-specific antibody, and two antibodies for detecting total actin. In addition, secondary reagents conjugated to DyLight® 488 or DyLight® 594 are included for labeling primary antibodies for detection using green (493Ex/518Em) or red (593Ex/618Em) filter sets.