By Jeffrey W. Pollard, John M. Walker
Read or Download Basic Cell Culture PDF
Similar molecular biology books
The second one version of Horizontal Gene move has been geared up to supply a concise and updated insurance of an important discoveries during this interesting box. Written by way of the main well known gene move and genome analytical scientists, this booklet info experimental facts for the phenomenon of horizontal gene move and discusses additional proof supplied via the new crowning glory of genomic sequences from Archea, micro organism, and Eucarya contributors.
This quantity plus its spouse, quantity 294, provides leading edge recommendations for the learn of ion channels, starting from the molecular to the physiological point. those volumes complement quantity 207 of equipment inEnzymology Key beneficial properties* meeting* Genetics* Electrophysiology* Expression platforms* version Simulations
The 1st quantity of the sequence, on "The balance of the Differentiated kingdom" bought many favorable reports from the clinical group. Many readers appear to believe us that e-book of topical volumes is a precious replacement to periodic compilations of particularly unrelated, notwithstanding updated stories.
- Essential Cell Biology, Second Edition
- Molecules and Life: An Introduction to Molecular Biology
- ECTO-NOX Proteins: Growth, Cancer, and Aging
- Counting Molecules Within Cells
- Signal Transduction Protocols
Extra info for Basic Cell Culture
Schmidt A, Hall A (2002) Guanine nucleotide exchange factors for Rho GTPases: turning on the switch. Genes Dev 16:1587–1609 29. Wolfman A, Macara IG (1990) A cytosolic protein catalyzes the release of GDP from p21ras. Science 248:67–69 30. West M, Kung HF, Kamata T (1990) A novel membrane factor stimulates guanine nucleotide exchange reaction of ras proteins. FEBS Lett 259:245–248 31. Yamamoto T, Kaibuchi K, Mizuno T et al (1990) Purification and characterization from bovine brain cytosol of proteins that regulate the GDP/GTP exchange reaction of smg p21s, ras p21-like GTP-binding proteins.
2 mg of Sulfo-SMCC crosslinker (Thermo Scientific). 6. Chemically synthesized HVR peptide (GKEKMSKDGKKK KKKSKC). 5 SortaseMediated Ligation 1. 15 μM sortase A (see Note 7). 2. 15 μM K-Ras catalytic domain stored at 4 °C. 3. Tris–EDTA buffer pH 9: 50 mM Tris–HCl, 10 mM EDTA, 10 mM MgCl2, 5 mM CaCl2 4. 10 mM βME. 5. 30 μM HVR peptide of K-Ras conjugated with S-farnesyl-Lcysteine methyl ester using Sulfo-SMCC crosslinker. 6 GTPase Activity Assay 1. 1 M GTP stored at −80 °C (see Note 8). 2. 10 μM fully modified K-Ras 4B stored at 4 °C.
1 Bacterial Expression 1. Place a single vial of one-shot BL21AI cells on ice and allow thawing. Add 1 μL of K-Ras 4B PET 42a plasmid to vial. Keep vial on ice for 30 min. 2. Heat the vial to 42 °C for 45 s and return to ice for a further 2 min. 3. After this, add 350 μL of sterile SOC medium to the cell vial and incubate with agitation at 37 °C and 250 rpm for 1 h. 4. Make a 350 mL solution of LB Broth supplemented with 350 μL of 50 mg/mL kanamycin solution. 22 μm pore-size membrane into a 1 L autoclaved conical flask.