Lab Exam 1 Flashcards
What is the relationship between water potential and solute concentration?
As the solute enters the cell, the water potential of the cell decreases relative to the outside environment (water moves from high potential to low)
What does a spectrophotometer measure in the cell membrane permeability lab?
It measured transmittance (percentage of light that was able to pass through the cuvette)
When you add solute to the cows blood, what happened in the cuvette?
It was first cloudy but then if the solute could pass through the membrane, the mixture would become transparent
Why did the mixture of cow’s blood and solute go from cloudy to transparent?
As solute, and water, entered the cell, it led to the red blood cells bursting (lyse)
What is hemolysis?
Red blood cells bursting
What is H50?
The time required to reach one-half the maximum value indicated by the plot
How do you calculate the time to reach H50?
(minimum transmittance % + (maximum transmittance % - minimum transmittance %))/2 = H50 transmittance %
Use the table given by spectrophotometer to find the time where it was closest to that value and then subtract that from the time the solute was first added
What is the relationship between H50 and membrane permeability?
Inverse relationship; the smaller the H50, the greater the membrane permeability to that substance
What is the relationship between membrane permeability and molecular weight for the water-soluble molecules?
Inversely related; as the molecular weight increases, it is less able to pass through the membrane
What effect does the net electrical charge on a molecule have on membrane permeability?
Membrane permeability for that molecule will be zero (solute is not able to pass through the membrane)
What solutions were not able to pass through the red blood cell membrane? Why?
Glucose (too big)
Sucrose (too big)
NaCl (charged ions)
How do lipid-soluble molecules pass through the red blood cell membrane?
They easily pass through because they dissolve in the lipids of the cell membrane
How does hydroxyl groups affect how a molecule passes through a membrane?
The more -OH groups a molecule has, the more water-soluble it will be and thus small uncharged water-soluble molecules can pretty easily pass through the membrane but for the larger molecules, it becomes more difficult
What is the relationship between lipid solubility and membrane permeability?
Directly related; the more lipid-soluble a molecule, the easier it can pass through the membrane
What does H50 stand for?
Hemolysis 50
What is diffusion?
The tendency of a solute to move from an area of high concentration to an area of low concentration
What are the three independent variables of the cell membrane lab?
- Size of solute molecule
- Net charge of solute particle
- Lipid solubility of solute molecule
What is the dependent variable of the cell membrane lab?
Cell membrane permeability
Why do you calibrate the spectrophotometer?
Using a blank solution takes into account the cuvette itself, thus, when you perform your other experiments, the cuvette will not absorb any light
Why is important to set a wavelength of maximum absorbance?
At this wavelength, even the slightest bit of change in concentration will cause a significant change in the absorbance of light
In the cell membrane lab, why was it important to first hit COLLECT on the iPad and then add the solute?
To be certain that we were recording everything that occurred when the solute was added
The cuvette had two sides with ridges and two sides that were clear, which side needed to be facing the light source?
The sides that were clear because the sides with ridges could affect the transmittance percentages, it may have absorbed some light which means that the data would be underestimating the actual amount of light that was being passed through
Why was important in the cell membrane lab to not have blood streaks down the side of the cuvette?
It could affect the time it took the red blood cells to lyse (for example it would take longer than normal) which then could affect the transmittance percentages and data values
What makes a molecule more lipid soluble?
Having more CH groups
What is the major property of proteins that influences their migration in an electric field?
The R groups of amino acids which generate a net electrical charge of a protein
Which type of amino acid R-groups is primarily responsible for this property?
Ionic category of R-groups
What are the properties of aspartic acid and glutamic acid?
They are negatively charged due to their carboxyl group when the environment surrounding them is at a high pH
What are the properties of lysine, arginine, and histidine?
They are positively charged at low pH environments because of their amino groups (R group)
What happens to aspartic acid when the pH changes?
At high pH environments, it will be negatively charged. However, as the pH decreases, the charge of aspartic acid will become more neutral because there are more protons (H+) to attach itself to the negative oxygens.
What happens to lysine when the pH changes?
At low pH environments, lysine has a net positive charge. However, as the pH increases, it will become more neutrally charged because more hydroxide ions will bond to the hydrogen (proton) atoms.
What is the typical trends of proteins at low pH environments and at high pH environments?
Typically, proteins have positive charges at low pH environments and negative charges at high pH environments
What is the isoelectric point of a protein?
The pH value in which the protein has a neutral (zero) net charge
What is the method used to separate and identify proteins?
Electrophoresis
What is the independent variable for the electrophoresis lab?
pH
What is the dependent variable of the electrophoresis lab?
The net electrical charge on each protein
What happens to aspartic acid as it enters a high pH environment?
Hydrogen atom that was bonded to oxygen in its carboxyl group bonds to the -OH ions that are in abundance, leaving oxygen with a negative charge, making the protein have a net negative charge
What happens to lysine as it enters a low pH environment?
A proton (H+) will bond to the nitrogen, giving it a positive charge, making the protein have a net positive charge
Which direction will a negatively charged protein move during electrophoresis?
Towards the cathode
Which direction will a positively charged protein move during electrophoresis?
Toward the anode
How can you isolate a specific protein from a group of proteins?
You can put the proteins in a pH environment that is the isoelectric point of the protein that you are trying to separate and applying positive and negative charges, which will filter all the other proteins but the one that you are trying to isolate
What does magnification mean in terms of a compound microscope?
Magnification enlarges the image of an object, making more things more visible
What is contrast in terms of a compound microscope?
Contrast uses light to differentiate between different parts of an object because different parts absorb varied amounts of light
What is resolution in terms of a compound microscope?
Resolution is the ability to distinguish two objects that are close together as being two separate objects
How much do the eyepieces of a microscope magnify?
Magnification is 10x