Immunoblotting is an effective approach that utilizes antibody-tags to analyze, isolate, and quantify proteins. A Western Blot, a type of immunoblotting method, was used in this experiment to evaluate the protein quantities contained within the different subcellular fractions. Although, the Western blotting method can be implemented for the study of more complex cellular processes. Complex cellular processes such as metabolism can be greatly evaluated with immunoblotting methods. For example, some
Beta-Galactosidase and Western Blot 0 1. EMG 9 and EMG 26 contain strain _lac-_(I- Z+ Y+) and strain _lac -_ (I+ Z- Y-)respectively.Three genes huddled together on the chromosome are required for two strains of _E.coli_ to utilize lactose.Consisting of three genes, namely, _lacZ_, _lacY_ and _lacA_, the _lac_ operon orderly handles these genes to code specific enzymes necessary for the metabolism of lactose. The genes _lacZ_, _lacY_ and _lacI_ would code for beta-galactosidase, galactosidase permease
Immunoprecipitation and western blot analysis LV-FLAG-HA-T2A-GFP and LV-hZNF804A-FLAG-HA-T2A-GFP constructs were transfected into HEK293 cells with N-terminal V5-tagged APEX1, EIF3K, FEZ1, LGALS1, PDHB, PDZD4, and RPSA overexpression constructs from DNASU. 72 h after the transfection, cells were lysed with cell lysis buffer (150 mM NaCl, 1.0% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris, pH 8.0) and collected by centrifugation at 14 kg at 4°C for 15 min. 3 mg of lysate protein was incubated
The technique used in this experiment was western blot to determine the protein levels in different cow’s stages; fetal calf serum, newborn calf serum and cow serum. Western blot is technique commonly used to identify proteins by its movement in the gel electrophoresis. Western blot is use to separate protein based on its molecular weight in gel electrophoresis. The proteins separated in the gel, then transferred to a nitrocellulose membrane using an electron current (1). Finally the membrane is
Proteomics is the study of protein and how they interact and function within organisms. This results in catalyzing and cataloguing proteins; while comparing how each variation of protein expression in different conditions. This is done in a large scale manner; a good example of this is the Human Genome Project that took place from 1990 to 2003. With the field of Proteomics it has given the ability to understand genes and to comprehend why there’s so much diversity of protein, including their importance
to localize the expression of a gene or its product (Northern, western, in situ hybridization, immunofluorescence, etc.), to determine the structure of the protein (NMR, crystallography ...) or to find partners proteins (double hybrid, immunoprecipitation ...) give important information on the function of a gene (V Ecochard -2011). This experiment will focus on SDS PAGE of purified protein and cell lysate samples, Western blot analysis procedure, interpretation of purified protein and lysate
Nematodes were cultured and handled at 20 °C using standard methods (Brenner, 1974). C. elegans strains used in this study are as follows: N2 Bristol and MR156: rrIs01[Pelt-2::GFP; unc-119(+)] X, which were used as wild type. CB3514: lin-23(e1883)/dpy-10(e128) II, YHS47: lin-23(e1883)/dpy-10(e128) II; rrIs01[Pelt-2::GFP + unc-119(+)] X, DH2: zyg-11(b2) II. DS97: mat-2(ax76) II, EU593: mel-26(or184) I, EU640: cul-2(or209) III, UP63: mat-3(ku233) III, NJ582: cul-1(e1756)/unc-69(e587) III, VC141: zif-1(gk117)
Western Blotting. Western blotting is the transfer of proteins from a gel to a membrane for subsequent analysis. This method can produce qualitative and semi-quantitative data regarding the desired protein. Western blotting requires multiple steps involving gel electrophoresis for separation of macromolecules, followed by blotting onto a matrix, selectively probed, and lastly detected using a technique dependent on the desired protein and experimental design.9 Kinase Assays. Kinase assays measure
Microtubules are an essential part of eukaryotic cells for various functions and processes, and are located throughout the cytoplasm in the cytoskeleton. Microtubules are essential for multiple cellular processes such as, cellular division and cellular movement and transport(6). Since microtubules are required for many cellular processes, mutations involving microtubules can cause various diseases including neurodegenerative diseases and cancer(5). One of the most common neurodegenerative diseases
decreased appetite, and losing weight, fever and complained of a sore throat with white patches developing. Her neck lymph nodes were also showing signs of swelling. Her Physician immediately ordered an Enzyme-Linked Immunosorbent (ELISA) test, Western Blot testing, CD4 T-cell count, Complete Blood Count (CBC), Erythrocyte Sedimentation Rate (ESR), and a p24 Antigen test and was asked to come follow up when the results where in. ________________________________________ ASSESSMENT Upon Jane’s follow