For an Immunoprecipitation experiment:The cellular extract contains a protein labeled with a fluorescent dye, which emits green fluorescence under UV light. Explain your observations in IP-1, IP-2, and IP-3 tubes by considering the interaction among antibody 1 (or antibody 3), the fluorescent-labeled protein, and the protein-A agarose beads. ProcedureLabel 3 centrifuge tubes as IP-1, IP-2, and IP-3 with your group name.1.2.3.4.5.6.7.8.9.10.11.Vortex the Resin (Protein-A) tube briefly. Using a wide bore tip (cut the tip with a blade ora pair of scissors), pipette 15 µl Resin (Protein-A) suspension to each labeled tube.Wash the Resin (Protein-A): Pipette 0.1 ml PBS to each tube. Centrifuge for 5 seconds at1,000 g. Carefully remove and discard the supernatant without disturbing the agarosebeads.Set up three tubes as in the following table:TubePBSCellular extractAntibody 1Antibody 3IP-1 IP-2IP-3280 μ. | 290 μ. | 280 μ.10 μl10 μ. 10 μ.10 μ.Close all tubes tightly. Incubate the tubes at 4°C for 60 min. Invert the tubes 3-4 timesevery 10 min.Label tubes for collecting the supernatants as "IP-1 SN", "IP-2 SN", and "IP-3 SNCentrifuge all tubes 5 seconds at 3,000 rpm. Carefully transfer the supernatants to thelabeled tubes without disturbing the agarose beads.Add 300 μl PBS to each tube. Vortex briefly or tap the tubes with fingers to suspend theagarose beads.Centrifuge all tubes 5 seconds at 3,000 rpm. Carefully remove and discard thesupernatant without disturbing the agarose beads.Repeat step 8 and step 9.Observe the pellets and supernatants under a UV transilluminator. IP-1IP-2 IP-31712PZSA201SASNIP-2IP-3supernatants IP-1 SupernatantsSupernatants

Immunoprecipitation is a technique used to isolate or detect the presence of a specific protein…

For an Immunoprecipitation experiment:The cellular extract contains a protein labeled with a fluorescent dye, which emits green fluorescence under UV light. Explain your observations in IP-1, IP-2, and IP-3 tubes by considering the interaction among antibody 1 (or antibody 3), the fluorescent-labeled protein, and the protein-A agarose beads.

For an Immunoprecipitation experiment:The cellular extract contains a protein labeled with a fluorescent dye, which emits green fluorescence under UV light. Explain your observations in IP-1, IP-2, and IP-3 tubes by considering the interaction among antibody 1 (or antibody 3), the fluorescent-labeled protein, and the protein-A agarose beads.

Essentials of Pharmacology for Health Professions

7th Edition

ISBN:9781305441620

Author:WOODROW

Publisher:WOODROW

Chapter5: Abbreviations And Systems Of Measurement

Section: Chapter Questions

Problem 11CRQ

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