Immunostaining was used to investigate the expression of the Fibrillin 1 gene in muscle fibres. Two antibodies were used, Antibody A binds to a muscle membrane protein that is unrelated to Fibrillin. Antibody B binds to the N terminus of Fibrillin 1, whilst Antibody C binds to the C terminus of Fibrillin 1. What is the best explanation for the staining pattern see in the following figure Antibody A Antibody B Antibody C
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- What internal references can be used in western blot of plasma membrane protein,expect Na-K ATPase??Why might it be necessary to include the 50 mL cultures in order to express protein? This question is referring to a lab experiment for protein expression. We streaked our plates. Put 50uL of two antibodies (AMP and CAM) into 50mL LB broth. Then took a colony and mixed it in the flask with LB broth and antibodies. The next step is to incubate the flasks overnight. Then add 500uL of AMP to 500mL of LB broth. We will add 25mL of the overnight culture to the 500mL LB broth. Hopefully this is enough detail to answer the question. Thank you!..In an experiment assay, sometimes there are several dilutions involved.0.5 mL of (Bovine serum albumin) BSA is pipetted into 0.5 mL of PhosphateBuffer Solution (PBS).Then 0.2 mL of the mixture is pipetted into 0.8 mL of PBS.What is the total dilution factor for this assay?
- To 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 antibodiesThis is an SDS-PAGE of an antibody purification sample with IgG seperated from a Bovine Calf serum. Would you be able to describe the bands that are appearing and why it appeared on the gel this way? As well, a western blot was done after the SDS-PAGE and the bands that appeared were at 50 kDa and 150 kDa for the IgG and two elution lanes. May you please explain?For an Immunoprecipitation experiment: What is the function of the IP-2 tube in this experiment?
- (1) Why the ratio of PE/GFP fluorescence is measured in the FACS experiment, instead of measuring only total PE fluorescence? (2) Explain the effect that each mutation causes in the function of MC4R, and how they can be linked to disease.For a immunoprecipitation experiment: By Observe the pellets and supernatants under UV light. Which tube contains green fluorescent protein?A variety of organic chelating ligands have been synthesized to tightly coordinate radioactive metal cations to identify and treat malig- nancies by coupling the metal complex to a polypeptide linker attached to a monoclonal antibody that binds specifically to a cell surface re- ceptor. One such metal complex is illustrated in the diagram on the right. The radioactive Cu-64 cation is tightly coordinated by the che- lating ligand that is, in turn, conjugated to a peptide linker attached to a monoclonal antibody. (The antibody is not shown in this diagram.) When coupled to a specifically designed monoclonal antibody, the complex binds specifically to somatostatin receptors that are ex- pressed on the surface of neuroendocrine tumors. Subsequently the entire complex with the receptor is internalized, i.e., passed into the cytoplasm, where the radioactive metal cation kills the malignant cell. 1. CH3 HN NHNH HN NHNH HN Но НО NH HN S-S HN Но NH NH HN -NH HN- H2N OH NH ) Write the three…
- 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 analysisIn the Western blot shown here, proteins were isolated from redblood cells and muscle cells from two different individuals. Oneindividual was unaffected, and the other suffered from a diseaseknown as thalassemia, which involves a defect in hemoglobin. Theblot was exposed to an antibody that recognizes β globin, whichis one of the polypeptides that constitute hemoglobin. Equal totalamounts of cellular proteins were added to each lane. Explain these results.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 signal