Immunostaining was used to investigate the expression of the Fibrillin 1 gene in muscle fibres. Twoantibodies 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 figureAntibody BAntibody AAntibody C A point mutation in the fibrillin gene introduced a stop codon in the proteinA gene translocation has removed the N terminus of the proteinA gene translocation has removed the chromosome encoding the fibrillin geneA tuncation mutation has removed the central region of the proteinThe C and N termini of the Fibrillin protein are unchanged by the mutationA point mutation in the fibrillin gene has changed an amino acid in the proteinA point mutation in the fibrillin gene has introduced a start codon in the proteinA truncation mutation has removed the N terminus of the proteinA point mutation in the fibrillin gene has resulted in the removal of a stop codonBoth the N and C termini regions of the protein have been deleted by a mutationA point mutation in the fibillingene has led to it not being expressed

Immunostaining is a process of staining proteins by antibodies that are covalently linked to an…

Answered: Immunostaining was used to investigate…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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Amino acid sequence analysis of all of the peptides found in a single IgG antibody would reveal unique peptide sequences totaling ~600–700 amino acids. Using this estimate, the predicted molecular weight of an antibody protein would be ~70–75 kDa. Yet, an intact antibody protein has a molecular weight of ~150 kDa. The explanation for this discrepancy is: IgG antibodies have many more heavy amino acids in them than most other proteins. Each IgG antibody is a complex of two identical light chains and two identical heavy chains. IgG antibodies tend to aggregate together during purification, thereby distorting molecular weight estimates. Each IgG antibody is a complex of four identical polypeptides. IgG antibodies are produced as dimers of two identical IgG monomers.
In the human immune system, so-called B cells canmake more than a billion different types of antibodymolecules that protect us from infection. However, ourgenomes have only three genes that encode the polypeptides found in antibodies. What experiments could youperform to determine what kind of combinatorial eventsoccur at the DNA level (V-D-J joining) and RNA level(alternative splicing) for any of these genes?
Regarding the production of recombinant monoclonal antibodies, it is CORRECT to state that: * (JUST ONE STATEMENT IS CORRECT) (A) Chimeric and humanized antibodies differ in relation to the percentage of sequence of the constant chain of molecules from the human genome (B) Chimeric antibodies are industrially produced by joining purified protein fragments of the Fab fraction of murine antibodies and the constant portion of human antibodies. (C) When using cells dependent on the DHF amplification system, we must use a vector containing the coding information of the light and heavy chains and another vector that contains the coding sequence of the enzyme dihydrofolate reductase (D)The industrial purification of monoclonal antibodies should preferably use an initial step based on the use of affinity columns, which will bind the antibodies by the Fc fraction (E) The use of amplification systems such as GS and DHFR has as main objective to make the production of monoclonal antibodies…
If you were to engineer some non-adhesive cells to express E-cadherin at either high levels or low levels and then mixed the E-cadherin high cells with the E-cadherin low cells, what do you predict will happen?
If you were to engineer some non-adhesive cells to express e-cadherin at either high levels or low levels and then mixed the e-cadherin high cells with the e-cadherin low cells what do you predict willl happen?
An important mechanism for generating diversity in immunoglobulin light chain V-region sequences is based on the fact that the RAG recombinase generates hairpin structures, rather than blunt ends, at the cleavage sites between the recombination signal sequences and the coding sequences. Explain how this mechanism generates diversity at the junctions.
In a tissue engineering experiment, stem cells are grown in a circular dish (diameter = 3 cm) with a thin layer of cell culture media covering the cells. It is expected that these cells will differentiate when exposed a new differentiation factor (small molecule) at a surface concentration of 50 ng/cm? During the experiment, 1 ug of the factor is placed at the center of the dish. Assume the diffusivity of the factor in cell culture media is 1 x 10^-5 cm^2/s. A. How long does it take before the cells 1 cm from the center of the dish begin to differentiate? B. Do you expect the cells at the edge of the dish differentiate? Justify your answer with calculations.
Given the following protein mixture, answer the following questiona. using hydrophobic interaction chromatography, what is the protein that will be eluted first|b. using affinity chromatography the protein that will be eluted LAST in an antigen containing column isc. the protein with the least migration in SDS-PAGE is?
In rare instances, B cells can be found that have two immunoglobulin light chain alleles, both of which are rearranged in frame, and can encode functional light chain proteins. Yet, on the surface of the B cell, only one of the two light chain proteins is detected in the membrane-bound immunoglobulin receptor. The reason these rare cells have two functional light chain rearrangements but only express one of the two light chains as part of the B-cell receptor is: One of the two light chains is formed from rearrangement of a V gene segment that is a pseudogene. One of the two light chain proteins doesn’t form a stable complex with the heavy chain expressed in this cell. One of the two light chain alleles is not transcribed efficiently, and produces only low levels of protein. One of the two light chain alleles uses a V gene segment that is not targeted very often by the RAG recombinase. One of the two light chains is rapidly degraded after synthesis due to improper folding.
You are investigating the activity of different cadherins in cells. You use recombinant DNA technology to express GFP-E-cadherin (green) in some L-cells (which do not normally contain cadherin) and YFP-P-cadherin (yellow) in other L-cells. Explain results you expect in three cultures containing: (1) only cells expressing GFP-E-cadherin, (2) only cells expressing YFP-P-cadherin, and (3) a mixture of GFP-E-cadherin-expressing and YFP-P-cadherin-expressing cells. Also predict how (a) knocking down the ability of the cells to synthesize fibronectin (FN) or (b) adding EGTA, which depletes free Ca2+ concentration in the culture medium, might affect the results.
According to the research article: “Telomerase Modulates Wnt Signalling by Association with Target Gene Chromatin,” how did the authors determine that TERT, Brg1, and beta catenin were all part of the same protein complex on TCF sites in vivo? Describe the assay used to determine this.
Describes a method known as Western blotting that can be used to detect a polypeptide that is translated from a particular mRNA. In this method, a particular polypeptide or protein is detected by an antibody that specifically recognizes a segment of its amino acid sequence. After the antibody binds to the polypeptide within a gel, a secondary antibody (which is labeled) is used to visualize the polypeptide as a dark band.For example, an antibody that recognizes α-galactosidase A couldbe used to specifically detect the amount of α-galactosidase A proteinon a gel. The enzyme α-galactosidase A is defective in individuals with Fabry disease, which shows an X-linked recessive pattern of inheritance. Amy, Nan, and Pete are siblings, and Pete has Fabry disease. Aileen, Jason, and Jerry are brothers and sister, and Jerry has Fabry disease. Amy, Nan, and Pete are not related to Aileen, Jason, and Jerry. Amy, Nan, and Aileen are concerned that they could be carriers of a defective…

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