||Recombinant Human CAPZA1&CAPZB(non-muscle isoform) with the alpha-1 SU untagged and beta-2 SU His-tagged was produced in HEK293 cells.
||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). Ca++-insensitive barbed end capping protein. Kd actin binding = >1nM (barbed end) .
||150mM NaCl, 20mM Hepes pH 7.4, 1mM DTT and 5% sucrose, when reconstituted with 1.0ml ultrapure water to obtain a 1.0 mg/ml solution.
||Capping activity: F-actin containing 20% pyrene actin 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 centigrade.
||>94% by scanning densitometry from Coomassie G-250 stained SDS-Gels.
||CapZ is stored at –70 centigrade 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 centigrade in glycerol or CryoProtect (Cat.#: 6011-01). Avoid repeated freeze / thaw cycles.
||At ambient temperature. Upon delivery store at -70 centigrade.