Anti-TCPTP抗体(ab1996)

Thank you for your enquiry and for filling the on-line Questionnaire. This antibody has been tested for Western blot application only, and we do not have any information if it is suitable for IP. Unfortunately, we do not have any information on IP. Ab1996 has been characterised for Western blot assay using recombinant TCPTP fusion protein. As the datasheet indicates it is expected to cross-react with Human (100% identity with immunogen) due to sequence homology. We would suggest running purified protein and applying positive control (i.e. human T cell or placenta) along with the samples. Do the endothelial cells express TCPTB at high or low levels? Do you have any preliminary background information? When you prepare the cell lysate, use RIPA buffer rather than standard NP-40 and 2X sample loading buffer which contain a cocktail of different proteinase inhibitors. It is important to use RIPA in order to prevent the degradation of the target protein in the samples. Preparation of RIPA cell lysates for Western Blots: 1. Wash cell pellets once with ice-cold PBS. 2. Add 1 ml of RIPA buffer to 10x8 cells, incubate on ice for 20 min, vortex 2 to 3 times. 3. Centrifuge for 5 min at 4oC at maximum speed in a microfuge tube. 4. Transfer supernatant into clean tube. Measure protein concentration with a protein assay. 5. Adjust concentration to 5 mg/ml with RIPA lysis buffer. 6. Add equal volume of 2 x SDS sample buffer into cell lysate, boil for 5 min. 7. Store at -20oC for daily use or -80oC for long term. Avoid repeated freeze thaw cycles. RIPA Base Ingredients Tris-HCl: 50 mM, pH 7.4 NP-40: 1% Na-deoxycholate: 0.25% NaCl: 150 mM EDTA: 1 mM PMSF: 1 mM Aprotinin, leupeptin, pepstatin: 1 microgram/ml each RIPA Protease Inhibitors Phenylmethylsulfonyl fluoride (PMSF) (200 mM stock solution in isopropanol; store at room temperature) EDTA (calcium chelator; 100 mM stock solution in H2O, pH 7.4) Leupeptin (store frozen in aliquots, 1 mg/ml in H2O) Aprotinin (store frozen in aliquots, 1 mg/ml in H2O) Pepstatin (store frozen in aliquots, 1 mg/ml in methanol) We hope this information will be useful for you.