Untitled Flashcards Set

What is immunofluorescence? - A technique to detect proteins using antibodies tagged with fluorescent dyes.

What is the role of secondary antibodies in immunofluorescence? - To bind primary antibodies and emit a detectable fluorescent signal.

What fluorophore color emits at 540 nm? - Green.

What is used to determine protein localization in mitochondria? - Antibodies against mitochondrial proteins and fluorescent secondary antibodies.

What is aerobic metabolism? - A process where pyruvate is converted to acetyl-CoA, enters the TCA cycle, and generates NADH and FADH2 for ATP production.

Where does the radioactive 14C in pyruvate go in aerobic metabolism? - Into molecules like acetyl-CoA, citrate, and downstream TCA cycle intermediates.

What is oxidative phosphorylation? - ATP generation in mitochondria using the electron transport chain and a proton gradient.

What is oxidized during oxidative phosphorylation? - NADH and FADH2 lose electrons, becoming NAD+ and FAD.

What does phosphorylation mean in oxidative phosphorylation? - The addition of a phosphate group to ADP to form ATP.

How can mutations reduce ATP production in mitochondria? - By impairing electron transport or disrupting the proton gradient.

What happens to the pH in chloroplasts during ATP synthesis experiments? - The stroma becomes basic (pH 8), while the thylakoid space remains acidic (pH 4).

How does a pH gradient drive ATP synthesis? - H+ ions flow through ATP synthase from the thylakoid space to the stroma, facilitating ADP phosphorylation.

What is the control in chloroplast ATP synthesis experiments? - Chloroplasts in an acidic solution with ADP and phosphate, showing minimal ATP production.

How are ATP generation processes similar in photosynthesis and oxidative phosphorylation? - Both use a proton gradient to drive ATP synthase.

Theory the cell is the smallest unit of all living things self reproducing enzymes has a membrane driven by free energy cells contain DNA into RNA into proteins in prokaryotes there are no nucleuses in eukaryotes there is a nucleus your transcription takes place when cells divide DNA is condensed and made visible how to study cells? Doctor hook in the 1600s discovered the cells so in my notes there is a chart that has a graph that shows how much you can see with each of these types of microscopy and so with the unaided eye you can see from 10 meters to 1mm so about human height to a frog egg with light microscopy you could zoom into a frog egg and see up to like 100 micrometers which shows organelles and some bacteria and then electron microscopy you can see viruses and proteins and it goes down to 1 nanometer electron microscopy has two different types transmission and scanning electron microscopy electron microscopy can see sub cellular structures harder than 10 nanometers and light microscopy can see cells in 200 nanometers or larger a light microscope uses refraction of light rays to magnify an object the three different parts are light source condenser lens objective lens and eyepiece lens you can magnify up to 110 or 1500 times and it resolves ups .2 micrometers the way it works is the condenser lens focuses light the objective lens magnifies and directs light into the eyepiece the ocular lens magnifies electron microscopy sprays a specimen with electrons a lenses are magnets that focus electrons beam bounces for scanning electron microscopy or passes through a very thin slice of the sample for transmission electron microscopy you stained the sample with heavy metals in the dark spots are electrons the sample is not alive magnification can be up to 2,000,000 times in the resolution goes as low as .2 nanometers scanning electron microscopy looks more 3D and is focused on the outside while electron transmission electron microscopy looks at the inside so here's a microscope flow chart that shows the two types of microscopy light and electron microscopy and then shows light microscopy shows cell tissues and is greater than 200 nanometers it can show live specimens which are called active progresses or static time points for dead specimens in phase contrast is a way to make like microscopy easier to see for electron microscopy yeah transmission electron microscopy which shows the inside of the cell and scanning electron microscopy which shows the outside of the cell which is for less than 200 nanometers for light microscopy shading or dyes can be added to the cells to show bright field shows a bright background ideal for colored specimen phase contrast heightened contrast produces a Halo effect and shows some transparent items differential contrasts interference gives a 3D quality and has no Halo active processes require a live sample fixed structures can analyze structure fluorescence labeled cells with fluorescence fluorophores can absorb at one light wavelength and emit a larger one a few things that are labeled with floor floors are antibodies in proteins so small synthetic molecules like fluorescent and rhodamine can be attached to antibodies GFP green fluorescence protein can be conjugated directly onto a protein of interest to show that protein fluorescence microscopy light is passed through two sets of filters one to excite the fluorophore and the other to show the wavelength immunofluorescence antibodies for antigens are tagged with secondary antibodies with fluorescence to show the location lower floors that recognize primary antibodies are less expensive because you can get them at different colors instead of primary antibodies that recognize antigens in multiple different colors which are about 400 apiece the epitope is the region on the antigen that the antibody recognizes the B cells generate antibodies that recognize a specific protein or antigen have small variable parts to fit different antigens placing antigens into animals makes an antibody and the secondary antibody is made at a different species floor floor protein is expressed via transgene tagged with the GFP and can be used to tag promoters for gene’

Which of the following is NOT a requirement of the cell theory to define how cells are the basic unit of life? - Ability to generate proteins from RNA
Size of bacterium - micrometers
Size of frog egg - millimeters
What molecule stores genetic information in cells? - DNA
Absorb photons and release a portion of the energy at a longer wavelength - fluorophore
Express recombinant protein tagged with a fluorescent protein - visualization of protein with a fluorophore
Use a primary antibody specific for a protein of interest - immunofluorescence
What approaches can be utilized to study the localization of protein-of-interest (POI) within a LIVING cell? - Transfection of cells with a recombinant plasmid containing the gene sequence for POI followed immediately by the gene sequence for GFP such that one fusion protein is produced upon expression
What approaches can be utilized to study the localization of protein-of-interest (POI) within a FIXED cell? - Transfection of cells (prior to fixation) with a recombinant plasmid containing the gene sequence for POI followed immediately by the gene sequence for GFP such that one fusion protein is produced upon expression
Living systems are incredibly diverse in size, shape, environment, and behavior. It is estimated that there are between 10 million and 100 million different species. Despite this wide variety of organisms, it remains difficult to define what it means to say something is alive. Which of the following can be described as the smallest living unit? - cell
Prokaryotic cells do not possess - a nucleus
Since cells are colorless, which of the following techniques allow visualization of cells in a microscope? - Cell stains
Which types of microscopy use an optical component to take advantage of different refractive indices of light passing through different regions of the cell? - phase-contrast
Which type of microscopy employs a light microscope and requires that samples be fixed and stained to reveal cellular details? - bright-field
Which type of microscopy requires the use of two sets of filters to observe the specimen? - fluorescence
Which type of microscopy has the ability to resolve cellular components as small as 2 nm? - transmission electron
Which type of microscopy requires coating a sample with a thin layer of a heavy metal to produce three-dimensional images of the sample surface? - scanning electron
How might a researcher study the presence of a specific protein in cells from different tissues of the body? - Treat cells with a dye that selectively binds to the protein of interest, and look for evidence of this dye in cells using light microscopy.
If you wanted to express GFP in the neurons of a fruit fly, which part of the expression vector would be MOST important to include in addition to the sequence for the GFP gene? - The DNA sequence for a neuron-specific promoter in fruit flies
The emission wavelength of the fluorophore is longer than that of the excitation since energy is lost as electrons move from the excited state to the ground state. - wavelength
Green fluorescent protein (GFP) and the gene that codes for the GFP protein: - can be expressed in eukaryotic cells
For each of the following sentences, match with the best word or phrase: - B: Secondary
Match each molecule with the metabolic process where it is FIRST metabolized or used in metabolism: - Glucose: Glycolysis
The addition of electrons to NAD+ to form NADH is a(n) - Reduction reaction; NADH is a(n) Electron carrier.
Photosynthesis involves the - Reduction of NADP+.
ATP is often used to add phosphate groups to proteins by kinases in a(n) - Phosphorylation reaction.
Which of the following is NOT a major part of glycolysis? - Electron transport
Match the following processes with either mitochondria, chloroplasts, or both: - Electron transport occurs: Both
Water is a source of electrons - Chloroplasts
Oxygen is produced - Chloroplasts
ATP synthase uses a proton gradient to phosphorylate ADP - Both
An inner membrane system is used to create a proton gradient for energy production - Both
NADH is a major electron carrier - Mitochondria
Oxygen accepts electrons - Mitochondria
Water is produced - Mitochondria
A proton gradient is generated by electron transport - Both