WebIf your vector has been cut with a single enzyme, you'll want to treat it with Calf Intestinal Alkaline Phosphatase (CIP) first. Set up the following reaction: * The amount of insert DNA used will depend on the relative size of the insert compared to the size of the vector. A molar ratio between insert and vector should be calculated. WebDocuments This purified calf intestinal alkaline phosphatase (CIP) is supplied in Tris buffer and 50% glycerol for use in protein research methods. The main applications for alkaline …

Protein dephosphorylation protocol Abcam

WebCIP (calf intestinal phosphatase) 10× CIP buffer (200 mM Tris-HCl, pH 8.0, 10 mM MgCl 2, 10 mM ZnCl 2) Water-bath or heating-block A. Phosphatase treatment with BAP 1 Prepare … WebThe phosphatase can be added directly into the digestion reaction during or after DNA digestion CIP is active in all NEB restriction enzyme buffers DNA purification is required … northern lights casino washington state

Protein dephosphorylation protocol Abcam

WebThe changes in activity and unfolding of calf intestinal alkaline phosphatase (CIP) during denaturation in different concentrations of trifluoroethanol (TFE) have been investigated by far-ultraviolet circular dichroism and fluorescence emission spectra. Unfolding and activation rate constants were measured and compared, the activation and inactivation … WebDephosphorylation by CIAP was carried out according to these protocols: From Reference 1: In a reaction mixture containing 50 mM Tris-Cl buffer, pH 7.5; 1 mM dithiothreitol; 5% glycerol; 5 mM MgCI 2; 0.1 to 0.2 mg/mL of phosphoprotein, and 10 U alkaline phosphatase. Web1 µl Calf Intestinal Phosphatase (CIP; 0.1 U/µl) Incubate 1 hr at 37°C. Remove the Calf Intestine Alkaline Phosphatase by use of the Phosphatase Removal Reagent (available in the KinaseMax™ Kit) or by the following protocol: To the above reaction, add: 125 µl water 15 µl 5.0 M ammonium acetate or 3.0 M sodium acetate 150 µl phenol/chloroform northern lights ccv login