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Bio 201 Lessons 5-7

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  • What is the correct binding order for Immunohistochemistry (IHC)?

    Tissue Antigen --> Primary Antibody --> Secondary Antibody --> Fluorescent Tag

  • What is the correct order in which cellular components will pellet when disrupted cell mixtures are centrifuged at increasingly higher speeds?

    Nucleus, mitochondria, ribosomes, proteins

  • True or False: Heavier cellular components will pellet first and lighter cellular components will pellet last

    True

  • What does the primary antibody bind to specifically in a Western Blot?

    The target protein or antigen

  • True or False: You can visualize a protein just by its size after running an SDS-PAGE

    False, there may be many proteins of the same size in a cell

  • What is the "blocking" step in a Western Blot?

    It prevents non-specific binding of antibodies to the nitrocellulose membrane

  • Where is the signal molecule typically bound when using an antibody for a Western Blot?

    To the secondary antibody at the Fc region

  • What side of the target protein or antigen does the primary antibody bind to?

    The Fab Region

  • Microscopy for visualizing cell components like the size of proteins, lipids, and ribosomes

    Electron Microscopy

  • What type of microscopy is better at visualizing samples that are thicker and more three dimensional?

    Confocal fluorescent microscope

  • True or False: The cells being visualized in brightfield stained and phase-contrast techniques must be fixed (dead)

    False

  • Immunohistochemistry visualizes __________ on a ___________

    Protein, microscopy slide

  • In Immunohistochemistry (IHC), which antibody binds first to the tissue?

    Primary Antibody

  • Technique used to detect specific proteins from a mixture of proteins typically from cell lysates

    SDS-PAGE/Western Blot

  • Technique used to detect specific proteins in a tissue (microscopy); can be both chemical and fluorescence

    Immunohistochemistry (IHC)

  • How are we able to start identifying a protein?

    By its size

  • In high concentrations of SDS, what happens to all the proteins?

    All proteins are converted to the same shape

  • In lower concentrations of SDS, what denatures proteins?

    Only the hydrophobic tails

  • Why can the negatively charged heads of SDS denature proteins?

    The heads cannot outcompete water for interactions with polar amino acids

  • If you performed Immunohistochemistry (IHC) on wildtype and mutant cells, what additional information could this give you?

    Location of the enzyme in the cells

  • If a competitive inhibitor was added for an enzyme in wildtype cells, what would you expect to see?

    The enzyme would be present but it is nonfunctional or less functional

  • What type of mutation occurs when a single codon is changed that causes the amino acid to change as well?

    Missense Mutation

  • What type of mutation occurs when a single codon changes but does not cause the amino acid to be changed?

    Silent Mutation

  • What is the relationship between the template strand and the mRNA/coding strand?

    They must be antiparallel and complementary to each other

  • What type of mutation occurs when a single codon changes and causes the amino acid to be changed into a stop codon?

    Nonsense Mutation

  • What type of mutation occurs when a codon is inserted or deleted that leads to a shortened and often non-functional protein?

    Frameshift Mutation

  • In Immunohistochemistry, the ______ region of a primary antibody binds to an antigen. The ____ region of the secondary antibody binds to _________

    Variable, variable, primary antibody

  • What toolbox technique is best suited to address this research question: How much insulin is present in a patient's blood?

    ELISA

  • What toolbox technique is best suited to address this research question: How does treatment with a drug affect protein Z's expression?

    Western Blot

  • What toolbox technique is best suited to address this research question: Is enzyme X expressed in a specific strain of yeast?

    Western Blot

  • What toolbox technique is best suited to address this research question: Is protein Y found exclusively in the mitochondria?

    Immunohistochemistry

  • What toolbox technique is best suited to address this research question: How to measure the amount of opioid in a person's urine?

    ELISA

  • What toolbox technique is best suited to address this research question: Which cells in a given tissue express the insulin receptor protein?

    Immunohistochemistry

  • What types of areas of SDS interact with the hydrophobic regions of the proteins?

    Hydrophobic

  • What types of interactions occur between the negatively charged heads of the SDS molecules and the positively charged R groups of proteins?

    Electrostatic Interactions

  • True or False: Smaller proteins migrate faster down the gel matrix

    True

  • What type of interactions lead to highly specific binding between an antibody and antigen at an epitope on the antigen?

    Weak Noncovalent Interactions

  • Multiple polypeptide chains that make up enzymes

    Subunits

  • True or False: Some antibodies only recognize a protein in its native folded form

    True

  • True or False: Some antibodies can still detect denatured forms of proteins

    True

  • Proteins that serves as a loading control; able to make a Western Blot quantitative and allows us to look at protein levels at different conditions relative to something common

    Housekeeping Protein

  • Technique used to measure the concentration of proteins, hormones, and other substances in fluid

    Enzyme-Linked Immunosorbent Assay (ELISA)

  • Cells only with receptors for a signal that can respond to that signal

    Target Cell

  • What does a signal's chemical property predict about the location of the receptor?

    Whether its receptors will be located on the cell surface or inside the cell

  • True or False: Proteins have an overall charge and are large molecules

    True

  • What is the flow of information for the central dogma?

    DNA --> RNA --> Protein

  • Why can we not go from the protein sequence to DNA sequence? (Why is the central dogma unidirectional?)

    Most amino acids are encoded by multiple proteins

  • Which molecules play a role in regulating the flow of information?

    RNA outside the protein coding region and DNA outside the transcribed region

  • True or False: mRNA is processed in bacteria but not in eukaryotes

    False, mRNA is processed in eukaryotes but not in bacteria

  • True or False: mRNA must exit the nucleus before translation can begin

    True

  • What is the molecular code needed to produce a precise product, determine the product's function, and faithfully reproduce that product

    Genetic Information

  • What are some key differences about bacterial genes that make it different in comparison to eukaryotic cells?

    Bacteria have smaller genes, no introns, and the gene length is the same as the mRNA length

  • What are some key differences about eukaryotic cells that make it different in comparison to bacteria cells?

    Eukaryotes have larger genes, typically multiple exons, and the gene length is longer than mRNA because introns are spliced out

  • The start of a gene that regulates transcription

    Promoter

  • Where does the ribosome begin translation?

    When it encounters the first AUG codon

  • True or False: The information coding for the start and stop of translation is part of a gene

    True

  • A stretch of DNA that codes for an RNA and maybe a polypeptide

    Gene

  • Two or more alternative sequences or forms of a gene

    Allele

  • A change in what region of a gene could lead to no transcription?

    Promoter

  • A change in what region might affect the tertiary structure of a protein?

    Between the start and stop codons

  • Why is yeast a good model organism for molecular work?

    The yeast genome has less non-coding regions than humans

  • True or False: A purine is a single ring structure

    False, a purine is a double ring structure

  • True or False: A pyrimidine is a single ring structure

    True

  • What nucleotide bases are purines?

    Adenine and Guanine

  • What nucleotide bases are pyrimidines?

    Cytosine, Thymine, and Uracil

  • DNA is a far more stable molecule than RNA. The difference in stability comes down to what carbon of the pentose sugar?

    The 2' carbon

  • What carbon does the nitrogenous base attach to?

    The 1' carbon

  • What carbon does the phosphate group attach to?

    The 5' carbon

  • What is the difference between ribose and deoxyribose?

    Ribose has a OH group at its 2' while deoxyribose is missing that oxygen and only has the hydrogen

  • What is the charge on phosphate?

    Negative

  • True or False: Nucleotides have a nitrogenous base, a five carbon sugar, and a phosphate group

    True

  • How many hydrogen bonds do A-T base pairs form?

    2

  • How many hydrogen bonds to G-C base pairs form?

    3

  • When a nucleic acid is synthesized, what direction does it grow from?

    5' to 3'

  • True or False: RNA can fold in ways that create unique structures/functions

    True