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Technical Data Sheet

AM0081

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β-Actin clone M008
Mouse Monoclonal IgG2b
Price
Size
Species Reactivity
MW

$295
100 μl
Hu
42 kDa

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Western blot analysis of human HUVEC-CS (lane 1), rabbit spleen fibroblast (lane 2), human Jurkat (lane 3), human LNCaP (lane 4), human HeLa (lane 5), and mouse F9 (lane 6) cell lysates. The blot was probed with mouse monoclonal anti-β-Actin (AM0081) at 1:1000 (lanes 1-6).

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Immunocytochemical labeling of β-Actin in paraformaldehyde fixed human MeWo cells. The cells were labeled with mouse monoclonal anti-β-Actin (clone M008). The antibody was detected using goat anti-mouse DyLight® 594.

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Application
Dilution


ELISA
1:1000


ICC
1:50


IP-MS
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
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.


Background 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.
Immunogen
Clone M008 was generated from a proprietary antigen related to human β-actin expressed in MDA-MB-231 breast cancer cell line.
Buffer and Storage
Mouse monoclonal, protein G purified antibody is supplied in 100 µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at -20°C. Stable for 1 year.
Specificity
Clone M008 detects a 42 kDa* protein corresponding to the molecular mass of β-actin on SDS-PAGE immunoblots of human cancer cell lines, as well as human recombinant β-actin. This actin antibody preferentially detects human β-actin with only weak reactivity toward actins in rat, mouse, or rabbit. The antibody works in multiple applications including western blot, immunocytochemical labeling, ELISA, and immunoprecipitation. In addition, mass spectrometry analysis of immunoprecipitates using AM0081 in human A431 cell lysate confirmed that this antibody detects β-actin.

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