By Simon M.I., Thorner J., Emr S.D.

The significantly acclaimed laboratory regular for greater than 40 years, tools in Enzymology is without doubt one of the so much hugely revered guides within the box of biochemistry. on account that 1955, every one quantity has been eagerly awaited, often consulted, and praised through researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence includes a lot fabric nonetheless proper this day - really a necessary booklet for researchers in all fields of lifestyles sciences.

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Calafat, H. Janssen, A. E. Von dem Borne, and A. Sonnenberg, Biochem. J. 336,153 (1998). lo A. D. Blagoveshchenskaya, E. W. Hewitt, and D. F. Cutler, J. Cell Biol. 145,1419 (1999). 3, for 21 hr at 35,000 rpm Abbreviations Tfn, transferrin; EGF, epidermal growth factor; NAGA, N-acetyl#-oglucosaminidase; SLMV, synaptic-like microvesicles; DCG, dense core granules. MD) is maintained in Dulbecco’s modified minimum Eagle’s medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS) and gentamicin (100 ,uglml).

Therefore, even if an AP fusion polypeptide is proteolytically cleaved, the fragments may remain associated in solution and may function perfectly well as a probe. Procedure for Immunoprecipitation Fusion Protein of Alkaline Phosphatase 1. Couple a monocolonal antibody against AP to Sepharose beads. a. 5 g of CNBr-Sepharose powder (Pharmacia, Piscataway, NJ). Swell the gel for a few minutes by mixing it with 1 mM HCl in a 50-ml tube. Wash the mixture in a sintered glass funnel over vacuum with about 500 ml of 1 mM HCl over a period of 15 min.

Wagner, Eur. J. Cell Biol. 75,331 (1998). 9P. W. Modderman, E. A. Beuling, L. A. Govers, J. Calafat, H. Janssen, A. E. Von dem Borne, and A. Sonnenberg, Biochem. J. 336,153 (1998). lo A. D. Blagoveshchenskaya, E. W. Hewitt, and D. F. Cutler, J. Cell Biol. 145,1419 (1999). 3, for 21 hr at 35,000 rpm Abbreviations Tfn, transferrin; EGF, epidermal growth factor; NAGA, N-acetyl#-oglucosaminidase; SLMV, synaptic-like microvesicles; DCG, dense core granules. MD) is maintained in Dulbecco’s modified minimum Eagle’s medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS) and gentamicin (100 ,uglml).

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