Microcentrifuges

Screening is usually carried out with liver microsomes from humans, rats, mice, dogs and monkeys and liver S9 fraction from aroclor 1254-induced rats. The incubation is typically mn with a volume of 0.2-1. OmL in a microcentrifuge or a glass tube. Different incubation conditions are used for CYP and UGT reactions. The incubation mixture for formation of oxidative metabolites and/or GSH conjugates contains ... 

Elution. Incubate the resin for 30 min with rolling in 500 /il Elution Buffer to competitively elute retained proteins. Collect the supernatant after centrifugation at 9300 xg for 5 min at 4° in a microcentrifuge (10,000 rpm in an Eppendorf 5415R). 

Mix by vortexing, then place the tube in a microcentrifuge and spin for 5 minutes at maximal rpm. 

Incubate specific mouse monoclonal primary antibody with FITC -conjugated mouse IgG specific Fab fragments at a ratio of 1 2 (weight for weight, based on concentration data supplied by manufacturers) in a small volume (e.g., in 10 pi or more, typically 1 pg of primary antibody in 10 pi) of staining buffer in a microcentrifuge tube for 20 30 min at room temperature. 

Using a micropipettor, add 10 pL of the reaction buffer to a small reaction vial or microcentrifuge tube. Add 15 pL of DNA. Then, with a fresh micropipet tip, add 15 pL of one enzyme. 

Mix all tubes by microcentrifuging for 5 min at 15,000 rpm, or by tapping tubes on any flat surface at least 50 times. Then incubate in a water bath at 37°C for 50 min. Carry these tubes over to Experiment 66. 

Prior to injection, the prepared sample should be centrifuged for 2 to 3 min in a microcentrifuge to remove any particulate material and to degas the solution. This practice is particularly important if the sample contains protein aggregates or other large particles, and when polymers are used to increase the viscosity of sample and ampholyte solution. 

A series of microcentrifuge tubes is set up, containing the membranes (usually 0.1-0.5 pM in receptor binding sites approximately 0.05-0.25 mg protein/ml) in Torpedo membrane protein and [ H]acetyl-choline concentrations ranging from 0 to 2.0 pM. Nonspecific binding is determined in the presence of excess unlabelled acetylcholine (100 pM-1 mM). 

Mix and transfer the contents of each microwell to a 1.5-ml microcentrifuge tube. 

Following DNase 1 digestion, the synthesized RNA is extracted with 1 volume of TE-saturated phenol/ chloroform, vortexed for 1 min and centrifuged in a microcentrifuge (12,000 x g) for 5 min. The aqueous phase is transferred to a fresh tube, 1 volume of chloroformusoamyl alcohol (24 1) is added, and the tube is vortexed for 1 min and centrifuged (12,000 x g) for 5 min. The aqueous phase is transferred into a fresh tube and cRNA precipitated by adding an equal volume of 5 M ammonium acetate and 2 volumes of ice-cold ethanol or an equal volume of isopropanol. The precipitation is carried on dry ice for a minimum of... 

To elute the cDNA synthesis probe, the spin column is inserted into a clean 1.5-mL microcentrifuge mbe 100 pL buffer NE are added to the column and soaked into it for 2 min. The column is centrifuged as described in step 9, with the flowthrough now containing the labeled cDNA probe. 

Brumback [28] has described a custom robotic system designed to automate the extraction of proteins from plant samples. Leaf or callus material (5—25 mg) is presented to the robot in microcentrifuge tubes, the system performs buffer dispensing, grinding, centrifugation, and pipetting unit operations, and a cleared supernatant is delivered in a 96-well microassay plate format for subsequent analysis. 

RIA Procedure. The RIA was developed using an antiserum pool which contained 2.0 mg/ml of anti-STXOL antibody. The protocol was designed for use in a 1.5 ml microcentrifuge tube. 

This assay was developed to be completely performed in a microcentrifuge tube. The basic procedure involves addition of a... 

Ligate vector and gene product To a microcentrifuge tube add ... 

Sterile microcentrifuge tubes Sterile pipettes and pipette tips SDS-polyacrylamIde gel electrophoresis system... 

A sterile plastic box that can accommodate six 100 ml tissue culture plates Microcentrifuge tubes Sterile pipette tips Microscope... 

Remove an agarose plug over the plaque using a sterile pipette tip and place in a microcentrifuge tube containing... 

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