Cell Biology

These questions must be answered by only using the power points provided and NCBI/PUBMED. It is very important that if you use any information from the powerpoints to cite by saying “According to Dr. Bush” or something along the lines of that. There is no need for any other form of citation like a specific slide and so on. Just make sure if you get it from the slides to give him credit. The power points and voice over info within them are to be the main source of info used for the assignment. If you need to use other sources outside of the PowerPoint (i.e. NCBI/PUBMED) then please officially cite them both in text and in a reference page. I assumed each question would need about a page for each question to be answered efficiently. If there is need for more then we can discuss as we go on how to compensate for that. It is important that each question has all details answered and covered.

 

Your lab has determined a section of the Rat Brain is involved in cheese foraging. Rats that have injury in that area do not eat or seek cheese snacks. That brain cell region is known as Get The Cheese (GTC). Each question 33 points in value.

Q1) Given your cell biology knowledge of cell-lines, detail a series of experiments to isolate a GTC cell line. Additionally, you must include your control experiments for the above techniques to confirm your experiments and results.  Also include a flow diagram of your techniques, experiments, and controls.

Q2) You next turn to GTC protein analysis. You are in search for GTC intracellular specific proteins that lead to the brain function in Rat’s GTC behavior Using your knowledge of protein purifications and 1D and 2D SDS PAGE techniques find specific GTC protein using other brain (OB) cell lines as controls to eliminate common brain specific proteins. Additionally, you must include your control experiments for the above techniques to confirm your experiments.  Also include a flow diagram of your techniques, experiments, and controls.

Q3) Using the protein purification techniques, proceed to generate monoclonal and polyclonal antibodies to GTC protein, sequence these GTC specific proteins via Edman degradation, and confirm usefulness of these antibodies by Western Blotting. Additionally, you must include your control experiments for the above techniques to confirm your experiments.  Also include a flow diagram of your techniques, experiments, and controls.

Q4) Next, you move to using flow cytometry and FACs (4 quadrants) to repurify GTC cells for primary cell culture testing confirming your results in part Q2 and Q3. Additionally, you use the Antibodies from Part Q3 to observe what intracellular organelles these proteins are in. Confirm by centrifugation, organelle fractionations and Western Blots.

You are expected to apply various centrifugation techniques including Differential/Density Gradient/Rate Zonal/Sedimentation Centrifugation. All your protein/organelle fractions will also be tracked by Spectrophotometric Analysis to track proteins within the subcellular fractions. Confirm your fractionations for purity, amounts of the Golgi/ER/Nuclear etc. membranes and proteins, and prove the fractionations worked.

Q5) After the experiments detailed in Question 4 you find the GTC brain cells have strange looking Golgi with 4 stacks instead of 3. Purify the GTC and OB cell specific Golgi organelles, protein fractionate using protein chromatography and HPLC to isolate just the Golgi proteins from both cell lines.

These Golgi proteins will then need to be visualized using 1D and 2D SDS-PAGE looking for differences in GTC vs OB cell lines. Western Blotting using polyclonal and or monoclonal antibodies will confirm protein levels and identity, protein similarity between the 2 cell-lines and protein differences.

1) Detail your experiments for first purifying the Golgi proteins, visualizing the Golgi proteins, and conformational experiments as described above.

2) Include all the needed techniques and control experiments as well

3) Include a flow diagram of your experiments using the techniques and controls.

4) Do the different tissues have different protein levels in their Golgi? Or the same. Discuss this key question!

Q6) Your lab now has GTC cell lines correctly labeled and analyzed, useful antibodies, pure GTC and normal brain Golgi fractions as well as a proteomic map of the key GTC Golgi proteins.

Given your knowledge of the Cell Free System first developed by the Classic Dr. William Balch Experiments which included Virus Glycoproteins, Mutant Cell Lines, Invitro Assays, Carbohydrates and Golgi Structure, design a series of experiments to develop a cell free system to test the hypothesis that the Golgi from the GTC and normal brain cell-lines to determine if they can interact and be both donor and acceptor Golgi elements (Cis-Golgi, Medial-Golgi, Extra GTC Golgi Stack and Trans-Golgi).

1) Discuss your experiments to test this hypothesis. You are free to use all techniques will be discussed as well as others if needed,

2) Included the essential controls experiments

3) Outline the series of experiments you use to set-up and demonstrate you have a successful cell-free system

Research Paper 101
Calculate your paper price
Pages (550 words)
Approximate price: -

Reasons to trust Research Paper 101

On Time Delivery

We pride ourselves in meeting the deadlines of our customers. We take your order, assign a writer but allow some more time for ourselves to edit the paper before delivering to you. You are guaranteed a flawless paper on a timely manner...

24x7 Customer Live Support

Our team at Research Paper 101 is committed to handling your paper according to the specfications and are available 24*7 for communication. Whenever you need a quick help, you can talk to our writers via the system messaging or contact support via live chat and we will deliver your message instantly.

Experienced Subject Experts

Online Experts from Research Paper 101 are qualified both academically and in their experiences. Many are Masters and Phd holders and therefore, are qualified to handle complex assignments that require critical thinking and analyses...

Customer Satisfaction

We offer dissertation papers as per students’ wishes. We also help craft out the best topics and design concept papers. By ordering with us, you are guaranteed of defending and making through those hard faced professors in the defense panel!

100% Plagiarism Free

We at Research Paper 101 take plagiarism as a serious offence. From the start, we train our writers to write all their papers from scratch. We also check if the papers have been cited appropriately. Our website also has a tool designed to check for plagiarism that has been made erroniusly. In essense, the paper you get will be 100% legit...

Affordable Prices

We understand that being a student is very challenging, some students balance between work and studies in order to survive. We therefore offer pocket friendly rates that are very competitive in the market.

Try it now!

Calculate the price of your order

Total price:
$0.00

How it works?

Follow these simple steps to get your paper done

Place your order

Fill in the order form and provide all details of your assignment.

Proceed with the payment

Choose the payment system that suits you most.

Receive the final file

Once your paper is ready, we will email it to you.

Our Services

No need to work on your paper at night. Sleep tight, we will cover your back. We offer all kinds of writing services.

Essays

Essay Writing Service

No matter what kind of academic paper you need and how urgent you need it, you are welcome to choose your academic level and the type of your paper at an affordable price. We take care of all your paper needs and give a 24/7 customer care support system.

Admissions

Admission Essays & Business Writing Help

An admission essay is an essay or other written statement by a candidate, often a potential student enrolling in a college, university, or graduate school. You can be rest assurred that through our service we will write the best admission essay for you.

Reviews

Editing Support

Our academic writers and editors make the necessary changes to your paper so that it is polished. We also format your document by correctly quoting the sources and creating reference lists in the formats APA, Harvard, MLA, Chicago / Turabian.

Reviews

Revision Support

If you think your paper could be improved, you can request a review. In this case, your paper will be checked by the writer or assigned to an editor. You can use this option as many times as you see fit. This is free because we want you to be completely satisfied with the service offered.