MCB Final Review

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6 Categories 30 Questions

•What was the overall purpose of the MCB Lab? What was your main hypothesis of your experiments?

Laura Sanchez-Michael

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Category 1

•What was the overall purpose of the MCB Lab?

To test the effects of potential anti-cancer compounds on human cancer cell lines using quantitative and qualitative assays.

What was your main hypothesis of your experiments?

Name and describe the compound you tested this semester

•What are positive and negative controls? What do they tell us? Why do we need them?

Positive control- A control known to produce an expected effectNegative control- A control that receives no treatment and should produce no effectThey tell us that our experiment is functioning correctly and we need them to verigy that our experimental data is trustable.

•What is a vehicle? What is it used for? Why do we use it as a control?

Vehicle control- Solvent that dissolves your compound; Used to confirm that your effects are caused by the drug itself and not the solvent.

Category 2

•What cell line did we use this semester? Describe it.

A549 non-small lung cancer cell line. This is an adherent cell line where cells stick to the bottom of the flask and require an enzyme (trypsin) to detach your cells.

What is cancer metastasis?

Cancer that spreads from where it started to a distant part of the body

•What is the difference between lymphatic and hematogeneous spread?

●Lymphatic spread refers to the spread of cancer cells through the lymphatic system, traveling via lymph vessels to nearby lymph nodes, while hematogenous spread refers to the spread of cancer cells through the bloodstream, reaching distant organs via blood vessels

Explain primary tumors

●Primary tumors develop at the site where the tumor first begins to grow. The most common sites for primary tumors are the skin, lungs, breasts, uterus, prostate, colon, and rectum.

Explain secondary (metastatic) tumors

•Cancer cells from a primary tumor can spread to other parts of the body, forming secondary tumors. This process is called metastasis. The secondary tumors are the same type of cancer as the primary tumor.

Category 3

•What environmental conditions do we incubate our cells in?

37 degrees Celsius; 5% CO2

What is a serial dilution?

When you dilute something step by step to make it less concentrated each time.

•Why is it important to remove the medium one row at a time?

It’s important to remove the medium one row at a time so the cells don’t dry out and die.

What is the range of the different micropipettes?

MULTIMEDIA

Explain qualitative vs quantitative data

Qualitative data describes what you see (words or pictures)Quantitative data measures what you can count (numbers)

Category 4

What were the 3 positive controls used throughout this semester?

H2O2, Ceramide, and Vinblastine

How does H2O2 kill the cancer cells?

Through Reactive Oxygen Species (ROS), which damages proteins, lipids, and DNA through oxidation

What does Ceramide do?

Blocks cell migration

What does Vinblastine do and how does this impact the cell cycle?

Blocks mitosis and causes the cells to arrest in the metaphase stage

What were the negative and vehicle controls used in your experiment?

Negative- DMEMVehicle- DMSO or Ethanol (Depending on your compound)

Category 5

What is the purpose of the protein titration curve in this lab?

To learn how to pipette accurately and to determine to concentration of an unknown protein sample

●What served as the standard protein for your curve?

Bovine Serum Albumin (BSA)

●What does it mean if your R² value is close to 1.0 when plotting your standard curve?

Your pipetting was accurate and consistent

What does BCA stand for?

Bicinchoninic Acid- The chemical that reacts with proteins to make color.

What is the difference between BSA and BCA?

BSA is the protein you measured and BCA is the chemical that makes it turn purple so we can measure it.

Category 6

What ion does BCA react with to produce color?

Copper ions

What wavelength is the BCA color change measured?

562 nm

What does a darker BCA color indicate?

The more protein in a sample

What 2 assays did you perform to assess Cell Viability at the EC50?

Crystal Violet Staining and In Cell Analyzer

Was the Crystal Violet Assay qualitative or quantitative?

Qualitative

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