How can we easily determine VEGF expression in a western blot experiment? By using fluorescent microscopy to view its transport into the cell By using a primary antibody targeting VEGF By adding purified VEGFR to an SDS polyacrylamide gel By doing a mass spectrometry analysis
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- How can we easily determine VEGF expression in a western blot experiment?
By using fluorescent microscopy to view its transport into the cell
By using a primary antibody targeting VEGF
By adding purified VEGFR to an SDS polyacrylamide gel
By doing a mass spectrometry analysis
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- Which of the following statements best describes the function of SDS in western blotting? Select one: A. It gives proteins a net positive charge. B. It can solidify polyacrylamide gel. C. It can form a colored product with proteins. D. It allows the transfer of proteins from polyacrylamide gel to nitrocellulose membrane. E. It can linearize protein molecules.The purpose for blocking the membrane for western blotting with milk is: to allow antibodies from binding to the membrane to allow antibodies to bind to the membrane to prevent the antibodies from randomly binding to the membrane to allow for the visualization of the signalYou have constructed a cDNA library in a phagemid vector.Describe how you would screen the library for a particulargene of interest. Describe methods using oligonucleotideand antibody probes.
- I need to make an antibody for the extracellular portion of the Notch receptor. I have made S35 labelled Delta and my boss has given me an antibody that recognizes Delta and is conjugated to a chromophore that gives off light. Place the following statements in order beginning with labeling. HINT: Labeling, protein enrichment, immune response, making monoclonal cell lines, antigen screening of monoclonal cell lines. A) Hybridoma cells are selected by plating them in HAT buffer. B) Using direct ELISA you coat individual tissue culture wells/dishes with a single monoclonal antibody. C) I detect my protein fraction using a geiger counter. D) You add a solution containing radiolabeled Notch extracellular domain to the wells. E) You only keep monoclonal hybridoma cell lines that give light. The other cell lines you throw away. You have your antibody! F) I centrifuge my homogenate in a sugar or salt gradient. G) I inject a mouse with my protein, giving booster shoots every week. H)…What are the advantages of the yeast-two-hybrid system over other in vitro methods used for protein-protein interactions? (select all that applies) enhanced sensitivity heavy dependence on antibody use no bias towards high affinity interactions precipitation of proteins of interestTo probe for presence tubulin in protein samples by western blot you use a anti tubulin monoclonal antibody. Which of the following secondary antibody should be used? one produced by a sheep immunized with goat antibodies. one produced by a donkey immunized with rabbit antibodies one produced by a goat immunized with mouse antibodies one produced by a frog immunized with hamster antibodies
- How would you determine the concentration of a protein that falls outside (above or below) the linear range of this assay? asapOne of the following statements is FALSE A. Labeled antibodies are the tagged probes of Southern blotting and ELISA (enzyme linked immunosorbent assay) B. In biopharmaceutical engineering; immunogenicity and pharmacokinetic properties are altered favorably; hence therapeutically C. In RFLP (restriction fragment length polymorphism), number and length of restrictionfragments vary due to genome polymorphism and bases sequencevariations D. In gene replacement therapy a normal gene allele substitutes permanently a mutant allele in a somatic cellWhich of the following pieces of information about our recombinant DHFR protein do you predict to obtain from Western Blot analysis? A) Protein size B) Protein sequence C) Protein function
- Sulfatase 1 (Sulf1) is a lizard blastema marker Identify a region of the Gekko japonicus Sulf1 protein sequence that is homologous with mouse Sulf1 (to educate antibody selection). hint: use NCBI Primer/Nucleotide BlastIn the bacterial transformation experiment, what is the primary purpose of using an ampicillin-containing medium? to select for transformed cells to select for wildtype cells to prevent contamination to stimulate a color changeYou are counting plaques on your plaque assay plates made from serial dilutions of your high titer lysate. Your 10-5 plate has 615 plaques although some are butting up against each other so it is difficult to get an accurate count. Your 10-6 plate has 42 plaques, and your 10-7 plate has only 1 plaque. Which plate would probably yield the most accurate titer calculation of your phage and why is it more trustworthy than the others?