In this chapter, we described co-immunoprecipitation as a method for identifying binding partners to a protein of interest. Asimpler 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 stepLast stepAnswer BankThe 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.

Co-immunoprecipitation (Co-IP) is a laboratory technique used to identify and characterize…

Answered: In this chapter, we described…

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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For a immunoprecipitation experiment: By Observe the pellets and supernatants under UV light. Which tube contains green fluorescent protein?
For an Immunoprecipitation experiment: What is the function of the IP-2 tube in this experiment?
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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 rhodamine
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Labs 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.
Describe three examples of how antibodies are used as reagents in cell biology and molecular biology.
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Our Western Blot experiment requires a buffer called TBS. At working concentration (1X), this buffer contains: 20 mM Tris 150 mM NaC1. Prior to use, however, the buffer is made and stored as a 10X concentrated stock What are the concentrations of these two ingredients in the 10X stock? Primary antibodies for the Western Blot experiment are prepared using this TBS buffer, with the addition of several other ingredients. Calculate the amount of each ingredient needed to produce10 ml of the final working antibody solution. b. 10X TBS stock c 0.01% (v/v) Tween detergent d. 3% (w/v) BSA (molecular weight 66430 g/mol) e. 1:10,000 dilution of antibody stock
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Give typed explanation

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