
Cap Z (capping protein)
Cat. #: 8322-01
(Actin Binding Protein)
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Description
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Protein
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Cap Z, Capping protein (non-muscle isoform)
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Origin
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recombinant, human
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Molecular mass
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66kDa
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Protein description
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The subunits of the heterodimeric capping protein CapZ possess a molecular mass of 34kDa (alpha 1-subunit) and 32kDa (beta 2-subunit, His-tagged). CapZ binds with high affinity in a Ca++-independent manner to the barbed end of actin filaments (1:1000 M/M).
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Actin interaction
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Ca++-insensitive barbed end capping protein. Kd actin binding = >1nM (barbed end) .
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Properties
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Form
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Lyophilized, ready-to-use.
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Quantity per unit
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2x 50 µg
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Buffer
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150mM NaCl, 20mM Hepes pH 7.4, 1mM DTT and 5% sucrose, when reconstituted with 50 µl ultrapure water to obtain a 1.0 mg/ml solution.
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Purity & Activity
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Purity: >94% by scanning densitometry from Coomassie G-250 stained SDS-Gels.
Capping activity: F-actin (Cat.#:8101) containing 20% pyrene actin (Cat.#: 8112) was incubated with CapZ and diluted below the critical concentration. Depolymerisation was followed by fluorometry and compared to an actin reference without CapZ.
Complete capping was obtained at a molar ratio of 1 : 1000 (CapZ : Actin). All proteins were used as supplied, i.e. without dialysis. G-Actin stock solutions were pre-spun 1h, 100.000xg, 4°C.
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Purification notes
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The alpha-1 SU is untagged, while the beta-2 SU contains a His-tag. Using anti-His antibodies, the protein can be identified by all suited immunological techniques.
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Protein concentration
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Determined by OD280 (0.1%=1.234).
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Storage instructions
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CapZ is stored at –70°C upon arrival will be stable in performance for at least 6 months from the date of purchase. The solubilized protein is kept on ice and should be stored at -20°C in glycerol orCryoProtect (Cat.#: 6011-01). Avoid repeated freeze / thaw cycles.
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Shipping conditions
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At ambient temperature. Upon delivery store at -70°C.
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Remarks
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For Use in Research only. Not for Use in Human or Veterinary Diagnostical or Therapeutical Applications.
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CAS no.
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Further Information
Product DataSheet
Material and Safety Data Sheet
Sequence on NCBI (alpha-1 SU)
Sequence on NCBI (beta-2 SU)
Structure on NCBI (PDB ID: 1IZN)
References
Myosin-II activity generates a dynamic steady state with continuous actin turnover in a minimal actin cortex
Sonal, Kristina A. Ganzinger, Sven K. Vogel, Jonas Mücksch, Philipp Blumhardt and Petra Schwille
bioRxiv preprint first posted online May. 2, 2018; doi: https://dx.doi.org/10.1101/312512
Yamashita A, Maeda K, Maeda Y
Embo J. (2003) 22 p.1529