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 cm2
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1. 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 cm2
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- 1) SDS PAGE gels can be probed with NBT/BCIP to perform an immunoblot true/ false 2) the direction of migration of protiens through a matrix is due to most proteins in natur having a net negative charge true/false 3) PBST is a buffer that is used for blocking the membrane during immunoblotting. true/false 4) the buffer used during for transferring proteins to nitrocellulose methanol. true/false 7) The size of the pores of a polyacrylamide gel dcreases as concentrations of polyacrylamide increases. True or false 10) The secondary antibody used in laboratory for western blotting is conjugated to which enzyme A) Alkaline decarboxylase B) GAPDH C) Alkaline Phosphatase D) Horseeradish peroxide 13) In protien electrophroresis, what reagent present in the sample buffer is used to eliminate difference in the charge densities of protein A) SDS B) 2- mercaptoethanol C) Tris-HCL pH6.8 D) Tris- HCL PH 8.8In 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 itTrue or false 1. Heterogenous antibodies used for structural studies can be obtained by monoclonal hybridoma technique. 2. PrP(Sc) is largely alpha-helical structure.
- 1. Antigens are made up of specific sequences of amino acids called ----- that determine their structure. A. epitopes B. multivalent sites C. paratropes D. constant regions 2. Which scenario best describes antibody cross-reactivity? A. A single pathogen with multiple epitopes is recognized by a multivalent antibody. B. Multipleepitopes are recognized by multiple antibodies C. A single epitope is recognized by multiple antibodies D. Two distinct, but structurally similar epitopes are recognized by the same antibody 3. What is the major benefit of cross-reactivity? A. Cross-reactivity permits the allergic response. B. Cross-reactivity promotes a faster immune response against unrelated pathogens. C. Cross-reactivity enhances immunity by providing protection against related pathogens. D. Cross-reactivity allows the immune system to respond to pathogens in various locations within the host (i.e., skin, mucosal tissues, extracellular fluids, etc.)Patient A, who is a recipient of USA-based poly X8650-CA vaccine for its autoimmune allergic rhinitis often visits the doctor twice a month to complain for “idiopathic colds”. Meanwhile Patient B, who is currently taking DOH pilot-used trial 1 mono E2128-PH vaccine for the same condition do not have to visit its doctor. Based on the principles of the antibody production, can you explain why patient A visits more often?Rheumatoid Factor RF Latex Test RF is a rapid latex agglutination test kit for the detection of rheumatoid factor (RF) in human serum. RF is found in sera of patients with rheumatoid arthritis and is believed to be IgM antibodies directed against the patients own immunoglobulin G. Principle: The RF latex particles are coated with specially purified human gamma globulin. When the latex suspension is mixed with serum containing elevated RF levels on a slide, clear agglutination is seen within 2 minutes. Questions 1. Is Rheumatoid Arthritis an autoimmune disease? 2. Differentiate the different types of Immunoglobulins 3. Differentiate Rheumatoid Arthritis from Gouty Arthritis
- ANTI - STREPTOLYSIN O TITER ASO Latex Test ASO is a rapid latex agglutination test for the detection of anti – streptolysin O (ASO) antibodies in human serum. ASO antibodies are found in sera of patients in response to infection with hemolytic streptococci of group A, C or G. Streptolysin O is highly antigenic therefore patients with these infections produce specific antibodies detectable by the reagent. Questions: 1. Illustrate how latex agglutination works to bind to ASO and form agglutinates 2. What is the significance of ASO? Is ASO specific to a disease? 3. What are Todd Units? Give the normal value of ASO1. True or False: Antibodies for Western blotting can be applied directly to the SDS-PAGE gel.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)…
- a) What is “washing” in immunology? What types of interactions doeswashing interfere with? What molecules are used for effective“washing” in the immunology lab?b) Blocking is also a common feature of immunological assays. Why isblocking required?i. A student forgot to block their Western blot membrane beforeadding the secondary antibody. They continue and block themembrane after the secondary antibody step, assuming itdoesn’t matter which order the steps are completed in. Howwould this affect the results seen?ii. A student blocked their ELISA plate with 50 microlitres ofblocking buffer, although their coating, binding and detectionsteps were at 100 microlitres. How would this affect the resultsseen?2. https://doi.org/10.1186/s12868-022-00692-1 (link to research) a) In the immunohistochemistry section of the materials and methods section the authors wrote “The number of positive cells in hotspot areas in ten high power fields (HPFs) in areas of demyelination and plaques in the brain stem were counted using the image analysis software (Lecia Application Suite Version 4.12.0, Welzlar, Germany).” Why were they looking at demyelination areas for this study? b) In the effect of mitoxantrone on histopathological changes in the brain section of the results section the authors wrote “Active plaques revealed inflammatory cellular infiltrates with abundant macrophages stuffed with myelin debris, an evidence of ongoing myelin breakdown.” What does macrophages stuffed with myelin debris have to do with the study?An MLS has completed an antibody panel using PEG. She cannot exclude anti-Fya, or anti-E. Anti-Kpª and anti-Ja cannot be excluded either but they are low incidence antigens and the MLS is not concerned with those at this time. What would be the next logical step to aid in antibody identification? a. Run the panel again using ficin treated cells b. Run the panel again using 22% albumin for enhancement c. Pay special attention to the strength of the agglutination reactions d. Run the panel again but increase the 37 degree incubation time to one hour