Your search for " University of California " resulted in 8 protocols
Β-Galactosidase Liquid Assay for Yeast Author: Ira Herskowitz Source:University of California. San Francisco Abstract: Grow the desired yeast strain to log phase in a liquid culture.
Measure the Optical Density at 600 nanometers (OD600) by diluting 0.5 ml of culture in 0.5 ml of media.
Place 1 ml of culture into a microcentrifuge tube .
Microcentrifuge at maximum speed for 5 min.
DNA-Dependent Protein Kinase Activity Assay Author: Donald Rio Source:University of California. Berkeley Abstract: 100 to 500 ng protein of protein substrate
1 μl of DNA-PK (HeLa Heparin 0.3 M pool)
1 μl of 10X DNA-PK Assay Buffer (50 mM final KCl concentration. see Hint #1)
1 μl of Excess DNA Solution
1 μl of γ-[32P]-ATP (10 μCi)
0.5 μl of 10 mM ATP
add ddH2O to give a final volume of 10 μl.
JNK/SAP Kinase Assay Author: Caroline Damsky Source:University of California. San Francisco Abstract: Grow cells to confluence in 100 mm culture dishes.
After cells reach confluence. grow in serum-free media for up to 48 hours.
Stimulate cells (see Hint #2).
Wash the cell monolayer with 3 ml of ice-cold 150 mM NaCl.
Add 1 ml of ice-cold Lysis Buffer.
Place cell culture dishes on a tabletop shaker at 4°C for 30 min.
Raf-1 Kinase Assay Author: Caroline Damsky Source:University of California. San Francisco Abstract: Place confluent cells in Serum-Free Media 48 hrs before the start of the experiment.
Stimulate the cells (the amount of time is dependent on the agonist and must be determined empirically).
To harvest. wash the monolayers with 3 ml of ice-cold 0.15 M NaCl and scrape into 1 ml of ice-cold Lysis Buffer.
Transfer the lysate to a 1.5 ml microcentrifuge tube and incubate on ice for 30 min.
Centrifuge 5 min at maximum speed in a microcentrifuge.
Transfer the cleared lysate to a new tube.
Removal of RNase from Bovine Serum Albumin By Acetylation Reaction Author: Michael Chamberlin Source:University of California. Berkeley Abstract: Add 2 g of Bovine Serum Albumin (BSA) to 100 ml of 50 mM NaHCO3.
Place the BSA solution on a stir plate with constant gentle stirring.
Place a pH probe in the solution and monitor the pH. Keep the pH at 8.0 by adding drop wise 1 M Na2HCO3.
Remove 600 μl of Acetic Anhydride from stock and place this in a beaker.
SLOWLY add Acetic Anhydride to the BSA solution. Do this drop wise by transferring a small amount of Acetic Anhydride into the BSA solution via a Pasteur pipette (see Hint #1).
Keep the pH at 8.0 by adding drop wise 1 M Na2HCO3.
RNase Protection and Immunoprecipitation of Nuclear Extracts Author: Donald Rio Source:University of California. Berkeley Abstract: Resuspend 60 mg of Protein A-Sepharose CL-4B or 375 μl Protein G-Sepharose in 7.5 ml of NET2.Add less than or equal to 0.3 to 20 μl of antiserum to 500 μl of Protein A-Sepharose CL-4b or Protein G-Sepharose in NET2.
Chloramphenicol Acetyl Transferase (CAT) Assay Author: Caroline Damsky Source:University of California. San Francisco Abstract: Wash a monolayer of cells three times using PBS Buffer.
Add between 0.5 to 1.5 ml of TBS Buffer and mix gently.
Incubate at room temperature for 5 min.
Scrape the cells to dislodge the cells from the culture plate and transfer the cell/TBS slurry to a 1.5 ml microcentrifuge tube.
Rinse the culture plate with an additional 0.5 ml of TBS Buffer and add to the microcentrifuge containing the cell/TBS slurry.
Centrifuge in a microcentrifuge for 2 min at setting "10" for 2 min.
Reverse Transcriptase Assay Optimized for Avian Sarcoma Virus Author: J. Michael Bishop Source:University of California. San Francisco Abstract: This protocol assays for reverse transcriptase activity by monitoring the incorporation of [3H]-dGTP into the DNA strand primed by the oligonucleotide dG primer and synthesized off a poly-C RNA template.
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