exam 1 Flashcards
this method breaks open cell membranes by disrupting lipid bilayers. It is usually used for mammalian cells. It is helpful for when you want to extract _____ or _____ from the cytoplasm
detergent, proteins, organelles
this method is used t beak cells apart using sheer force. It is commonly used with tissue samples, plant cells, bacterial cells or fungi/yeast. The mechanical force disrupts the cells which is important for cells with strong cell walls. It is common for large-scale ______ disruption
mechanical force, tissue
this method is used t beak cells apart using sheer force. It is commonly used with tissue samples, plant cells, bacterial cells or fungi/yeast. The mechanical force disrupts the cells which is important for cells with strong cell walls. It is common for large-scale ______ disruption
mechanical force, tissue
this method uses ultrasonic waves to create shear forces and cavitation bubbles that break open cells. It is effective for bacterial cells, mammalian or yeast cells. It disrupts both soft and ____ cells and is used for ____-_____ extractions or to target specific ______. (great for breaking open gram-positive or negative bacteria)
high frequency sound, tough, small-scale, organelles
high frequency sound may be used on yeast cells to extract proteins or ____ _____
nucleic acids
this method is used when you want to preserve the integrity of intracellular components (like _____ or ______) after cell lysis. It is popular for bacterial or mammalian cells and can be scaled for large _____
high pressure, proteins, enzymes, volumes
if a protein is not _____ correctly during a western blot, the epitope for the ____ _____ may not be excessible resulting in a weak or no signal
denatured, primary antibody
what location would a transcription factor be found in an adult organism?
nucleus
this method uses an antibody that is conjugated to a fluorophor.
co-localization
co-localization - step 1)
1. ___ cells obtained from the control and cells obained from the experimental group. ____ the cells so that the antibody can access ______ proteins. This antibody should be conjugated to a _______
fix, permeabilize, intracellular, fluorophore
co-localization - step 2)
2. express a _____ that encodes a nuclear protein fused to a fluorescent protein OR stain the cells with a ______ ______ conjugated to a different color fluorophore
transgene, second antibody
co-localization - step 3)
3. visualize cells under a _____ _____
fluorescent microscope
____ focuses on the location of one protein or molecule of interst where as ________ focuses on spatial overlap between signals from 2 or more proteins.
IF, co-localization
___ uses a primary antibody that binds to the _____ of interest, and a secondary antibody which is conjugated to a fluorescent dye that binds to the ___ ____
IF, protein, primary antibody
IF can be used to detect and visualize the presence of a ____ or _____ in a cell or tissue.
protein, antigen
t or f) lipids can be tested using BiFC
false, proteins
_____ test protein-protein interactions. It only works with proteins because it involves altering the ___ ____ that yield those proteins
BiFC, genetic sequences
t or f) lipids aren’t encoded by genes
true
Which model would be helpful to know the specific location of a certain amino acid in a protein that is known to be directly involved in the protein’s function
wire
after homogenizing and centrifuging cells, Joe extracts the _______, incubates the proteins with ___ and runs them in electrophoresis with _______ gel.
supernatant, SDS, polyacrylamide
After running ____-______ Joe transfers proteins to a nitrocellulose membrane because this membrane is more ______. This is part of the process for western blot
SDS-PAGE, durable
ribosome __ antibody
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these give proteins their flexibility and help them fold
intrinsically disordered sequences
(IF / direct / indirect) is a quantitative assay used to measure the concentration of a specific ANTIGEN by detecting its binding to a PRIMARY ANTIBODY that is directly conjugated to an enzyme
direct
direct ELISA:
1. the ________ (protein of interest) is coated
2. a ___ ______ which is directly conjugated to an ______ binds to the ______
3. a _______ is added which catalyzes a color change
4. intensity of color is proportional to the amount of ______ present in the sample
antigen, primary antibody, enzyme, antigen, substrate, antigen
indirect ELISA:
1. the _____ is coated on a microplate
2. the ___ ____ binds to the ______
3. a ___ ____ is conjugated to an ______ that binds to the ____ _____
4. a substrate is added to generate a color change
this directs the presence of __________ in a sample
antigen, primary antibody, antigen, secondary antibody, enzyme, primary antibody, ANTIBODY
direct ELISA detects _____/____ whereas indirect ELISA detects _________
antigens/proteins, antibodies
direct ELISA measures the ______ of a specific antigen where as indirect ELISA measure concentration with _______ of the signal via secondary antibodies
concentration, amplification
Loops, turns and beta sheets are all examples of residues that help maintain ______. an example is proline
rigidity
