PhosphoaminoacidanalysisofnonTCAprecipitableproteins
Phosphoamino acid analysis of non-TCA precipitable proteins1. Cut out gel slice, incubate 60 min with 1 ml 5.7 M HCl @ 110oC in 13 x 100 mm screw cap tubes.2. Add 4 ml water, centrifuge @ 2000 rpm in bench top centrifuge for 10 min, transfer supernatant, freeze and lyophilise.3. Dissolve residue in 10 ml water, add 0.4 ml 50% (v/v) slurry of Dowex AG 1 X 8 (Cl- form), adjust pH to 7.5 - 8.5 with NH4OH.4. Add 1 µl of ......阅读全文
Phosphoamino－acid－analysis－of－nonTCA－precipitable－proteins
Phosphoamino acid analysis of non-TCA precipitable proteins1. Cut out gel slice, incubate 60 min with 1 ml 5.7 M HCl @ 110oC in 13 x 100 mm screw cap
蛋白质检测
· Protein detection (Aberdeen's Lab)The method used to locate the proteins following 2D-PAGE depends on the nature of the original sample.
Phosphoamino－acid－analysis:Mark－Kampss－method
Phosphoamino acid analysis:Mark Kamps's method1. Label your protein with 32Pi. Then subject the protein to SDS polyacrylamide gel electrophoresis
蛋白质磷酸化
Tyrosine Kinase Assay Using Synthetic Peptides (T. Miller)Small synthetic peptide substrates are especially well suited for applications such as assay
蛋白质定量
Quantitative Determination of Peptides by Sulfhydryl (-SH) Groups New (Contributed by David Van Horn, Dept. of Chemistry, UC Berkeley Greg Bulaj, Dept
Preparation－of－Bacterial－Proteins－for－Analysis－by－2DPAGE
The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was princ
Analysis－of－Proteins－using－Small－Format－2D－Gel－Electrophoresis
Preparation of protein samplesIntracellular virus proteinsThe following method has been developed principally for the analysis of intracellular protei
磷酸氨基酸分析的方法特点和功能
中文名称磷酸氨基酸分析英文名称phosphoamino acid analysis定 义一种分析蛋白质中是否含羟基氨基酸(如丝氨酸、苏氨酸、酪氨酸等)-O-磷酸酯的方法。应用学科生物化学与分子生物学（一级学科），方法与技术（二级学科）
Amino－acid－composition
There has been a recent revival of interest in the use of AA composition for the identification of proteins from 2-D gels. This technique uses the idi
Purification－of－MBP－(maLTosebinding－proteins)－Fused－Proteins
Express fusion proteins as per the GST-fused protocol up to Step 7 (Day 3). All steps in protein purification should be done at 4° C unless otherwise
Arachidonic－Acid－Labeling
1) Grow cells to a density of 5-8 X 105 cells/ml in RPMI 1640 containing serum.2) Pellet cells and wash 1 time with room temperature PBS.3) Resuspend
The－Citric－Acid－Cycle
The Krebs cycle, also called the citric acid cycle, is a fundamental metabolic pathway involving eight enzymes essential for energy production through
Neutralizing－Arachidonic－Acid
NOTE - Use arachidonic acid from Biomol (# FA-003)1) Make up the appropriate concentration of AA in 100% ethanol.2) Add 1µl of phenol red solution.-->
Glycosphingolipid－analysis
1) Incubate cells with 1 µCi/ml of 3H-galactose for 72 hours.---> If treatment is for an extended period of time: treat in serum free media containing
Lipid－analysis
Thin layer chromatography is based on the separation of a mixture of compounds as it migrates with the help of a suitable solvent through a thin layer
Principles－of－nucleic－acid－hybridization
Principles of nucleic acid hybridization5.2.1. Nucleic acid hybridization is a method for identifying closely related nucleic acid molecules within tw
Oxidation－of－Polyunsaturated－Fatty－Acid
Unsaturated fatty acids are those that contain one or more double bonds in their alkyl-chain. Polyunsaturated fats with two double bonds usually have
Preparation－of－nucleic－acid－probes
Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s
DAPI－Nucleic－Acid－Stain
实验概要The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding
蛋白质翻译后修饰的验证问题
Why are proteins, detected by mass spectrometry, not validated by site-specific antibodies?The modified motif could be detected by mass spectrometry (
Synaptic－Proteins－at－the－Synaptic－Junction
The postsynaptic density (PSD) is a submembranous structure at the postsynaptic membrane mainly at the excitatory synapses. The neurotransmitter recep
Purification－of－GST－Fused－Proteins
Day 1Set up an overnight culture in 100 ml LMM broth or 100 ml terrific broth containing 100ul 100 mg/mlAmpDay 2Add 40-50 ml o/n culture to 1 lt terri
Analysis－of－Heme－and－Hemoproteins
Heme is perhaps the most ubiquitous cofactor found in nature and the most functionally diverse. Hemoproteins are involved in cell respiration (cyt
Flow－Cytometry－Analysis
PurposeFlow cytometry employs instrumentation that scans single cells flowing past excitation sources in a liquid medium. The technology can provide r
Lineage－Analysis－of－Blood
Materials:Capillary tubes1.5 mL Eppendorf microfuge tubes15 mL conical centrifuge tubes96-well V-bottom plates (Corning Costar 3894, from Fisher)Flow
Biosynthesis－and－Analysis－of－Bilins
The term bilin is a collective one to describe a broad group of open chain tetrapyrroles and derives from the name “bile pigments” as the first of
Analysis－of－Oligosaccharide－Ligands
Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity ChromatographyAnalysis of Oligosaccharide Ligands by High Performance Liquid A
CELL－CYCLE－ANALYSIS
PROPIDIUM IODIDE: The most commonly used dye for DNA content/cell cycle analysis is PROPIDIUM IODIDE (PI). It can be used to stain whole cells or isol
Analysis－of－murine－BM
Histologic analysis of murine BM is a necessary complement to flow cytometric or in vitro analysis. Techniques to do this are well established in huma
An－Introduction－to－Biocomputing
An Introduction to BiocomputingVersion 2.01October 1996David Steffen, Ph.D.President, Biomedical Computing, Inc.6626 WestchesterHouston, Texas 77005US