Homogenization buffer

Translation-competent ER membrane fractions can also be prepared from tissue culture cells. We recommend a terminally differentiated secretory suspension cell line, such as a plasmacytoma (e.g., J558L), which contains abundant levels of ER membrane. In this protocol, cells are collected by centrifugation (5 min, 500 x g) and resuspended in a homogenization buffer containing 10 mMKOAc, 10 mMK-HEPES, pH 7.5, 1.5 mMMg(OAc)2,... 

These hydrolytic profiles have been obtained mostly in homogeneous buffered aqueous solutions, but the natural water body contains many kinds of dissolved and suspended matters such as humic substances, metal oxides, and clay particles. The distribution of nonionic pyrethroid molecules in these matters can be explained in terms of a partition model. The association with these matters reduces the fraction of... 

The frozen infected silkworm pupae are homogenized in homogenization buffer. 

Muscle homogenate 1 400 in homogenization buffer. Because of the fast inactivation of the enzyme in the homogenate, assay the enzyme at four different concentrations immediately after homogenization and determine the protein concentration later on. The enzyme is stable in muscle at -20°C for about 7 days, and at -70°C for about... 

Homogenate buffer sucrose 171 mg/2 ml Tris-HCl/EDTA buffer (MW 342.30 —> 250 mM). Prepare fresh ... 

Make up two assay samples, one containing substrate mix with G-6-P [ 100 pi substrate mix+ 25 pi sample (homogenate) + 25 pi homogenate buffer = 150 pi] and one containing substrate mix without G-6-P [100 pi substrate mix+ 50 pi sample... 

Blank reaction without cell lysate mix 25 pi of homogenization buffer, 74 pi of reaction buffer and 1 pi of 100 mM GTP and proceed by starting the incubation for 1 h at 37°C in the dark, as described for the reaction mixture. 

Homogenizing buffer, 0.02 M tricine, pH 8, containing 0.01 M NaCl and 0.4 M sucrose. Keep cold. 

Fresh spinach leaves, about 50 g. These should be washed well with cold water, placed in plastic bags, and kept cold for several hours before use. Homogenizing buffer, 0.02 M tncine, pH 8, containmg 0.01 TWNaCl and 0.4 M sucrose. Keep cold. 

Diced beef muscle trimmed of visible fat and connective tissue Homogenization buffer (10 mM Tris Cl/1 mM EDTA, pH 8.0), 4°C Sodium hydroxide Ammonium sulfate... 

Homogenize 150 g diced beef muscle in a blender with 450 ml of homogenization buffer for 1 to 2 min at high speed. 

Buffer capacities for four heterogeneous systems are shown in Figure 6. For comparison, two homogeneous buffer capacities are also shown... 

The Revelle factor is about 10 for typical surface seawater. The details of the chemistry of this general relationship and its derivation have also been discussed by Sundquist et al. (1979), who called it the "homogeneous buffer" factor. Of interest is the fact that using the Revelle factor one can calculate for an instantaneous change in the Pco2 °f the atmosphere, the distribution of carbon between the atmosphere and seawater. 

Sundquist E.T., Plummer L.N. and Wigley T.M.L. (1979) Carbon dioxide in the surface ocean The homogeneous buffer factor. Science 204,1203-1204. 

Why is the tissue suspended in 0.2 M sucrose, at pH 7.2 Sucrose is used in the homogenization buffer to create a medium that is isotonic with the organelles. This prevents diffusion of water into the organelles, causing them to swell, burst, and spill their contents. A pH of 7.2 helps to decrease the activity of lysosomal enzymes and maintain the native structure of proteins in the sample. 

Homogenization buffer, pH 7.4. Concentrations given below indicate the final concentrations. The concentrations of stock solutions that can be used are indicated. 50 mM HEPES (stock solution 1M), 1 mM EDTA 0.5 M (stock solution pH 8.0), 10 mM MgCl2 (stock solution 1M), Protease inhibitor cocktail tablets (see Note 2), 2 iM Pepstatin A (stock solution 1 mM in 100% DMSO), 1 mM phenylmethyl-sulfonyl fluoride (PMSF) (stock solution 50 mM in 100% alcohol) (see Note 2). 

Resuspension buffer Same as homogenization buffer but without Pepstatin A and PMSF. 

Suspend the cells in cold homogenization buffer at 2 x 108 cells/mL. 

We use 5 mM L-histidine monohydrochloride (pH 7.0) as a homogenization buffer. For plant tissues containing secondary metabolites that can inhibit enzyme activity (e.g., resins, phenolics, tannins), complex extraction buffers are usually required.11 14,17 Examples of extraction buffers for... 

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