Documents about Polyacrylamide Gel

  • 5 Pages

    CELL BIO Lecture1_2009slides

    University of Texas, CH 53170

    Excerpt: ... icroscopy Optical Sections visualized by Confocal Fluor. Microscopy Alberts, 9-21 Alberts, 9-22 How to get things into cells Light vs. Electron Microscope Alberts, 9-34 Alberts, 9-42 Alberts, 9-45 Alberts, 9-50 4 SDS- Polyacrylamide gel electrophoresis (SDS-PAGE) Alberts, 8-19 Alberts, 8-17 SDS- Polyacrylamide gel electrophoresis (SDS-PAGE) Alberts, 8-18 Alberts, 8-18 Western Blotting Alberts, 8-20 5 ...

  • 3 Pages

    Lec20

    UC Riverside, BCH 120

    Excerpt: ... e receptors) also heterodimerize. Heterodimerization with a partner that lacks a DNA-binding domain (e.g. Id inhibitor binds MyoD). Dominant-negative effect of the defective partner (i.e. Id). Analysis of protein - DNA interactions in vitro 1. DNase I Footprinting DNA fragment to be analyzed is radiolabeled at one end with 32P (red dot at 5-end). A. DNase I in limiting amounts cuts radiolabeled DNA about once per DNA molecule randomly (arrows). B. If a protein sits on a specific DNA sequence, DNase I will not cut the DNA fragment at those nucleotide positions. C. The cut DNA fragments generated by Dnase I are then analyzed by electrophoresis on a denaturing polyacrylamide gel . This allows to map the location on the DNA of the protein binding site after autoradiography of the gel (visualization of a footprint). C. The cut DNA fragments generated by Dnase I are then analyzed by electrophoresis on a denaturing polyacrylamide gel . Samples containing the DNA-binding protein Undigested DN ...

  • 25 Pages

    Lecture 4 (Sept 5)

    Vanderbilt, BSCI 110B

    Excerpt: ... are proteins studied? Analyzing complex protein mixtures by SDS-PAGE. 1. "Denature" proteins by boiling in SDS and 2-ME 2. Load protein samples to crosslinked polyacrylamide gel 3. Apply strong electric field 4. SDS-coated proteins migrate towards positive pole. Smaller proteins move faster Small proteins can readily pass Larger proteins retarded by cross-linked polymer How are proteins studied? Analyzing complex protein mixtures by SDS-PAGE. 1. "Denature" proteins by boiling in SDS and 2-ME 2. Load protein samples to crosslinked polyacrylamide gel 3. Apply strong electric field 4. SDS-coated proteins migrate towards positive pole 5. Stain with protein dye or antibodies How are proteins studied? Analyzing complex protein mixtures by SDS-PAGE. How are proteins studied? X-rays Diffraction Determining the 3-D structure of a protein by X-ray crystallography. Protein crystal See Fig. 2.33 Karp 3-D protein structure Next lecture. The enzymatic functio ...

  • 21 Pages

    lec01a.ppt [Compatibility Mode]

    UCLA, LIFESCI LS 3

    Excerpt: ... ques commonly used in molecular biology labs Learn how to use bioinformatics tools and databases Answer various questions in biology How many different polypeptides are found in a protein and what are their sizes? How do living organisms respond to a change in environment? How can DNA be used to determine how related species or individuals are? Lab Hemoglobin: 2 !, 2 " ! Lab I: SDS gel electrophoresis " " Use polyacrylamide gel electrophoresis to determine number and size of subunits in a protein Basic technique essential in molecular biology and biochemistry How can we detemine the size and number of polypeptides in a protein? hemoglobin ! Purify protein ! Di Dissociate into separate polypeptides i t i t t l tid (detergent: SDS) ! Separate using polyacrylamide gel electrophoresis SDS Polyacrylamide gel forms matrix that separates p proteins based on size Lab 1 ! You'll be given unknown protein of g given MW " " " Denature Separate subunits on polyacrylamide gel Determine how many of each subu ...

  • 2 Pages

    BIO 320 Lecture1notes_2009

    University of Texas, BIO 50160

    Excerpt: ... BIO320CELLBIOLOGYSpring2009 T.OHalloran&A.DeLozanne Lecture1 MethodsinCellBiology(January20,2009) Reading:Alberts,5thEd.Chapter8,thefollowingsections: Isolatingcellsandgrowingtheminculture Cellscanbegrowninculture Purifyingproteins Cellscanbesep ...

  • 82 Pages

    Electrophoresis

    Georgia Tech, CHEM 4581

    Excerpt: ... tech nical manual protein electrophoresis Protein Electrophoresis technical manual tm 80-6013-88/Rev. B0/12-99 page nder Page nder This technical manual contains information that has been distilled from more than 30 years of laboratory experience in developing and verifying applications and product performance for protein electrophoresis. As such it is ideal for both new and current users of protein electrophoresis as both a teaching and a reference guide. The rst chapter provides a theoretical framework, and Chapters 2 through 4 cover the major aspects of protein electrophoretic separation: polyacrylamide gel electrophoresis, isoelectric focusing, and gel analysis. In addition, an extensive reference list is summarized at the end of every chapter, and a detailed glossary is included at the end of the manual. Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iii Chapter 1 Introduction to Electrophoretic Theory . . . . . . . ...