Why do we want to avoid making bubbles while working with liquids in the sterile hood?2.
What’s the first thing you do when setting up a laminar flow hood for sterile techniques?
The amount (usually expressed as a percentage) of growth surface covered by cells is called?4.
What’s the name of one microscope we are using today?
What should you have ready every time you come to lab?6.
Why do we use a phase microscope?7.
Why do we use trypan blue?8.
Carbon dioxide levels for cells are?9.
Equation for Newbauer and Rosenthal cell counting traysQUIZ 21.
What stain was used to determine cell viability?2.
Five squares from a hemocytometer were counted: live cells counted: 43,49,40,38,41. Dead cells counted: 1, 3, 1, 2, 1. What is the % viability?3.
List three conditions that must be taken into consideration when growing mammalian cells in an artificial environment?4.
During which phase does exponential growth occur in the eukaryotic growth curve?5.
A cell line that can be sub cultured indefinitely is called?6.
What enzyme are we using to remove adherent cells from their flasks?7.
Using counting conventions discussed in lab how many live cells are in this 1mm square?QUIZ 31.
What happens when cells can’t attach to vessel?
Why do we expect multicell tumor spheroids to form on agarose-coated dishes?3.
Using an antibody with an attached fluorescent molecule as a stain is called?4.
Name two chemical fixaters5.
Name one of the two filters that are necessary for fluorescent microscopy and explain (briefly) what is does.6.
There are 38, 43, 46, 40, 49 live cells per square using the Fuchs-Rosenthal hemocytometer. The cells have been diluted 1:2 with PBS and the total volume was 4.9ml. calculatecells/ml and total cell number. Show work.7.
What does the abbreviation MTS stand for?8.
Briefly describe the process of attachment for a cell in culture9.
List two nuclear stains10.
List two mitochondrial stains11.
List a liposomal stainQUIZ 41.
Fluorescent staining done in conjunction with an antibody is called?2.
Concanavalin A (Con A) is highly specific for what two sugars?3.
The purpose of treating cells with chemical fixatives is 3-fold. Name one.4.
What is a lectin?5.
Immuno-fluorescence attached to the protein of interest is what method?6.
Con A is part of a group of sugar-binding proteins known as?7.
You want to subculture some cells into a final volume of 3ml of medium with a final cell concentration of 2x10
cells/ml. if you have a suspension of cells at 1.5x10
cells/ml,how much of the cell suspension will you need? How much medium will you need?QUIZ 51.
Con A is made up of how many subunits? What is the MW of each subunit?2.
What fluorescent dye was conjugated to Con A for the experiment we did last week?3.
List one way you can elute bound protein from an affinity column.4.
What two sugars were used for the competition part of the Con A cell staining experiment?5.
What was the purpose of the well with no added stain for the experiment from Lab 5?6.
Why did we treat the cells with methanol (in Lab 5) before adding our fluorescent stain?7.
What do we know about the biological function of lectins?8.
What substance are we purifying Con A from?QUIZ 61.
When the crude extract is applied to the affinity column, what is the fraction called that passes through the column?2.
Why were we using sephadex
with no alterations as our chromatography matrix? Shouldn’t we have attached an antibody or some other protein to it?
What was the NMWCO of the ultrafilter we used? Why did we choose filters with that NMWCO?4.
How will we test the purity of the Con A sample?5.
How did we determine the biological activity of Con A?6.
Why is everything dissolved in 1X Con A buffer for hemagglutination assay?7.
Con A was extracted from what substance?8.
Protein measurements are taken at what absorbance when using a spectrophotometer?9.
What was the process called when forming dialysis using centrifugation ultrafilters?QUIZ 71.
Why do we have to add 10X Con A buffer to our protein solution?2.
Using a serial dilution of our Con A solution will let us determine the…?
Name two advantages of using SDS-PAGE4.
Differential centrifugation allows us to isolate?5.
Your Con A sample is at the concentration of 21.3 mg/ml. how much of your Con A sample will you need to prepare a sample at 2mg/ml in a 1X Con A buffer at a final volume of 400 microliters?6.
What method will be used to determine the purity/integrity of your pooled fractions (eluted fractions) of Con A?7.
Explain the results seen in wells with agglutination and wells without agglutination in the 96-well plate (what was happening to the RBC in each case and why?)QUIZ 81.
The tissue we will be using today is from?2.
List two of the components of the loading dye and give their purpose?3.
What type of matrix did we use for our electrophoresis?4.
How will we know when to stop the electrophoresis?5.
Which will have a smaller Rf 15kD or 65kD?QUIZ 91.
Which fraction should contain the highest quantity of mitochondria?2.
Which fraction should contain the highest quantity of nuclei3.
How did we homogenize the tissue in our experiment last week?4.
What two centrifugation methods are available for cell fractionation?5.
Why do we keep all tubes on ice?6.