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- You are interested in performing indirect immunofluorescence light microscopy to observe the localization of the catalase enzyme in the cultured HeLa cells, obtained historically from the cervical tumor of Henrietta Lacks. You were going through the lab stock and found a few primary and secondary antibodies. Which of the following secondary antibody can you use in your experiments? O All of the mentioned antibodies can be used in the experiment Goat anti-human antibody conjugated to 10 nm gold Goat-anti-human catalase conjugated to 10 nm gold O Human anti-catalase antibody conjugated to fluorescent rhodamine Goat anti-human antibody conjugated to fluorescent rhodamineIn immunofluorescence experiments, what is the role of the primary and secondary antibodies? (select all that applies) Group of answer choices The primary antibody binds the protein of interest whereas the secondary antibody binds the primary The primary and secondary antibodies can be interchangeable The secondary antibody is labeled with a fluorophore and is therefore responsible for generating the signal The secondary antibody is less important and therefore the experiment can be performed without itLabs that do a lot of Western blots often have hundreds of primary bodies but only two or three secondary antibodies. Why? I understand that secondary antibody aids in the detection, sorting or purification of target antigens by binding to the primary antibody which directly binds to the target antigen. However, I don't undrstand why there are only two or three secondary antibodies available in the laboratory. Glad if the expert would advise.
- 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 antibodies1. knowing that the Control is 6.22 x 10-20 moles.channel-2sec-1 and CFTR-CRISPR: 3.84x 10-20 moles.channel-2.sec how did they get the answer and explain how? Immunostaining using Quantum Dot fluorescence reveals that there are 5 CFTR channels/μm2 in the “CONTROL” and 81 CFTR channels/μm2 in the “CFTR-CRISPR”. Calculate the number of moles of Cl-transported per channel per second for both treatments. Note that there are 1x108 μm2 in each cm2Many ELISAs that are used to identify the presence of antibody to viruses have a 3rd control (along with negative and positive). In between the rows of wells coated with antigen, are rows of cells coated with tissue culture used to grow the virus. In other words, patient serum is added to wells that contain antigen along with wells that contain tissue culture. Why do you think this might be necessary?
- How does a rapid test detecting RSV work and what kind of binding does it use (competitve, sandwich, indirect)? Explain using anti-rsv antibody conjugated with gold nanoparticle, anti-rsv antibody, and anti-human IgG. Draw a diagram if possible.5) The immunoblot shows probing of samples taken from a time course after treatment with the proteasome inhibitor MG132. Recall that the proteasome is responsible for selectively degrading only those proteins that have been tagged with ubiquitin. The left indicates the antibody that is used to detect protein for each panel. P53 P27 AGR2 GAPDH MG132 (h) 0 0.5 1 2 4 8 12 24 Image from Wang et al., (2019) Oncogene 38:3458-3474. a) What effect does MG132 have on each protein? What does this tell you about each of these proteins? ● P53 P27 AGR2 GAPDH b) How do we know that the differences we observe in the proteins is not due to un- equal loading of samples into each lane of the gel?You have 4 ml of an antigen solution, how would you prepare a 3-fold dilution series such that you will have at least 2 ml of each dilution? Please draw a picture and explain, I don't understand what it means by 3-fold. Please explain, no copy paste asap
- The scale bar in panels (a) and (d) in this image is 50 um (micrometei. Question: Which of the answers below is equivalent to 50 um. Figure description: An immunofluorescence image of uninfected cells (top panel a-c) and virus infected cells (bottom panel d-f). a,d) show Hoechst staining (blue) of the nuclei and a merged image of the respective panels: E staining for the virus (red); and c,f) show staining for calreticulin (green). The scale bar (white line) represents 50 um. Hoechst/Merge Virus Calreticulin a C d e O50m 50 x 10 m 50 x 10m 50 x 10 m + Virus Uninfected cellsIf you wanted to make an assay detecting SARS-CoV-2 virus presence, like the assay shown in the previous question, what would be a logical combination of things to include in your list of supplies to obtain? rabbit anti-SARS-cov2 spike antibody, anti-rabbit IgG horseradish peroxidase-conjugated antibody, tetramethylbenzidine rabbit anti-SARS-cov2 spike antibody, anti-mouse horseradish peroxidase conjugated antibody, tetramethylbenzidine rabbit anti-SARS-cov2 spike antibody, anti-rabbit SARS-cov2 spike antibody, tetramethylbenzidine SARS-cov2 spike, anti-rabbit IgG horseradish peroxidase-conjugated antibody, tetramethylbenzidine rabbit anti-SARS-cov2 spike horseradish peroxidase conjugated antibody, anti-rabbit horseradish peroxidase conjugated antibody, tetramethylbenzidineIn this chapter, we described co-immunoprecipitation as a method for identifying binding partners to a protein of interest. A simpler variation of this method can also be used to isolate proteins of low abundance in a complex mixture. Arrange the steps in sequential order to use this technique for this purpose. First step Last step Answer Bank The protein mixture is centrifuged, the supernatant is removed, and the pellet is washed. The antibody binds to the protein of interest. The antibody-protein complexes become insoluble. An antibody against the protein of interest is added to the mixture. Protein A beads are added to the mixture.