You are trying to purify a protein that is soluble in a solution of 2 M ammonium sulfate. After centrifugation to remove other proteins that have precipitated at this high salt concentration, you recover the supernatant to assay the target protein's activity in a cell culture system.
(a) Explain why the cell dies when incubated with the supernatant.
(b) What procedure could you preform to correct the problem?

Answers

Answer 1

Answer:

a) The cell dies because it was in a hypertonic medium, and it loses water due to the difference in pressure and may even die from dehydration.

b) To correct this, the cells can first be placed in an isotonic solution (one that has the same concentration of solute as a solution separated or isolated by a semi-permeable barrier). A method to remove the salt is called dialysis.

Explanation:

A protein purification is a series of processes that allow the isolation of protein from mixture. Thereby this process is vital for the characterization of the function, structure and interactions of the protein of interest. It is therefore a very important procedure to perform correctly.

Ammonium sulfate is a salt whose chemical formula is (NH4)2SO4. The cells are in a 2M solution (2 moles of sulfuric acid per liter of solution). This is detrimental because the solution is hypertonic, it has a higher osmolarity in the external medium than inside the cell, so that a cell in such a solution loses water due to the pressure difference, i.e. osmotic pressure, and may even die from dehydration. The outflow of water from the cell continues until the osmotic pressure of the external medium and the cell are equal.

Centrifugation is a method by which solids can be separated from liquids of different density by means of a rotating force. After centrifugation, the supernatant will have a soluble part (proteins) and a pellet (cells and other insoluble parts). Thereby, the protein of interest would be recovered in the supernatant. But since the cell was not previously in a suitable medium, it could not carry out its metabolism and died and could not produce the protein of interest.

a) The cell dies because it was in a hypertonic medium, and it loses water due to the difference in pressure and may even die from dehydration.

b) To correct this, the cells can first be placed in an isotonic solution (one that has the same concentration of solute as a solution separated or isolated by a semi-permeable barrier). A method to remove the salt is called dialysis.

Dialysis is the process of separating molecules in a solution by the difference in their diffusion rates or osmotic pressure across a semipermeable membrane. A solution of various types of molecules is placed in a semi-permeable dialysis bag, such as, for example, on a cellulose membrane with pores, and the bag is sealed. The sealed bag is placed in a beaker with pure water or another solution. Molecules small enough to pass through the pores (often water, salts and other small molecules) tend to move in or out of the dialysis bag in the direction of the lower concentration. However, larger molecules (such as DNA, proteins or polysaccharides) that are larger than the pore diameter are retained within the dialysis bag.


Related Questions

What are the consequences of humans adding gases and other substances to earth's atmosphere?

Answers

Answer:

there is an increase of the average global temperature due to the accumulation of polluting gases in higher atmospheric levels that causes regional and global climate change.

Explanation:

It causes climate change,global warming. This is when the bubble around the earth deteriorates causing its biome to suffer.

population of zebras experienced ten births, six deaths to predation, three deaths to
disease, three individuals
pining the population, and zero individuals leaving. What is happening to the population growth?
It is increasing.
N
It is decreasing.
It is staying the same.
It reaches carrying capacity.

Answers

It is increasing Answer A

If there are more lynx than rabbit populations can support, how can there still be large lynx populations during that time?

Answers

Answer:

reputation hope this helps

How would expect the biomass to change after ten days if seeds and plants were provided the following conditions?
No light, water (seeds sprouted): Stay the same, decrease, or increase
Light, water (seeds sprouted): Stay the same, decrease, or increase
Light, no water (seeds did not sprout): Stay the same, decrease, or increase

Answers

Answer:

La biomasa (energía) se refiere a la biomasa «útil» en términos energéticos formales: las plantas transforman la energía radiante del Sol en energía química a través de la fotosíntesis, y parte de esa energía química queda almacenada en forma de materia orgánica; la energía química de la biomasa puede recuperarse quemándola directamente o transformándola en combustible.

Explanation:

Please select the word from the list that best fits the definition

Grouping items together to keep observations organized


inferring


predicting


classifying


observing

Answers

The answer is classifying.

~~~~~~~~~~~~~~~~~~~~~~~~~~~

Definition: Arranging (a group of people or things) in classes or categories according to shared qualities or characteristics.

Part of an important cellular process involving a DNA strand is modeled below.



What is the purpose of this cellular process?

Answers

Answer: Cellular respiration is the process by which cells in plants and animals break down sugar and turn it into energy, which is then used to perform work at the cellular level.

Explanation: In simpler words, it provides energy for the cell to do basic needs.

Rocky's class recently got pet turtles. When putting together the habitat, decomposers were added to the soil, as well as some smaller organisms that turtles eat. Some students wanted to get "fake" plants for the terrarium, but Rocky insisted that this would negatively affect the carbon and oxygen cycles.

Which of the following reasons support why Rocky is correct?

Group of answer choices

A) Carbon won't be removed from the air.

B) Photosynthesis won't occur.

C) The turtles won't be able to breathe.

D) All of these are correct.

Will mark as brainliest if correct

Answers

Answer:

The correct answer is - D. All of these are correct.

Explanation:

It is clear that the terrarium is sealed or not, if it is open to the outside environment or not. Considering it sealed and no outer interaction, then if a fake plant to this terrarium then there will be no photosynthesis will take place as there are no green plants to capture the sunlight and make glucose and no way to release oxygen for the other organisms.

If in this no green plants then the removal of CO2 will not be possible. Higher amount of carbon and without oxygen turtles would not be able to survive even though there is food for them.

A scientist was given some cells in a test tube .When she did karyotyping studies on these cells, she found that they each consisted of 23 chromosomes with only one sex chromosome in each. Half of these cells had one" X" chromosome in each of the cells and the other half had one"Y" chromosome in each of the cells. From this information, which would be a reasonable conclusion?
a.
These could be cells taken from any part of the human body
b.
These are all gametes
c.
There is no way to tell what type of cells they are.

Answers

Answer:

These could be cells taken from any part of the human body

what is the botanical name of beans​

Answers

Answer: The common botanical name for beans is Phaseolus vulgaris.

Answer:

Vhaseolus Vulgaris

Explanation:

The botanical name of bean is Phaseolus Vulgaris

4. How should the fossils in the sediment layers of Earth appear if species have evolved over time?
A. All of the layers should contain fossils of simple organisms.
B. All of the layers should contain fossils of complex organisms.
C. The lower layers should contain fossils of more complex organisms.
D. The upper layers should contain fossils of more complex organisms.

Answers

Answer:

D the upper layers

Explanation:

at first, lifeforms were simple. they got more complex for a variety of reasons, including to adapt to the changing earth. simple organisms will be lower down for that reason.

The lower layers should contain fossils of more complex organisms. So, the correct option is (C).

What are Fossils?

A fossil is defined as the preserved remains, imprint or trace of something that once lived from a previous geological era. Examples of fossils are bones, shells, exoskeletons, fossilized traces of animals or microbes, objects preserved in amber, hair, petrified wood, and DNA residues. The totality of fossils is known as the fossil record.

There are four main types of fossils which are formed in different way  are conducive to preserving different types of organisms. Fossils are mold fossils, cast fossils, trace fossils and true form fossils.

Fossil layers are fossils that formed in sedimentary rock that formed in layers by depositing and pressing sediments on top of each other.

Thus, the lower layers should contain fossils of more complex organisms. So, the correct option is (C).

Learn more about Fossils, here:

https://brainly.com/question/19486615

#SPJ6

You are studying aging in budding yeast. With each division, mother yeast cells bud and divide, forming a daughter that has renewed replicative potential (in other words, it is rejuvenated). The mother, however, does not show a renewed replicative potential. You find that the mother cell dies after a few more divisions, but the daughter cell dies after it divides 30 or more times. What would you expect to see at a cellular level

Answers

Answer:

hi my names emily and i support charli addison and the vegan teacher btw everyone go vegan

Explanation:

Ophrys sphegodes, the early spider orchid, has flowers with yellow-green sepals and petals and one highly modified, velvety brown central petal at the base of the flower. There is single male anther is found at the tip of the column. The stigma is on the underside of the column just below the anther.

Required:
a. Is the flower complete or incomplete?
b. Is the flower perfect or imperfect?
c. Is the flower radially or bilaterally symmetrical?

Answers

Answer and Explanation:

a. This is a complete flower. Complete flowers are those formed by chalice, corolla, androecium and gynoecium. In the case of the flower presented above, we can see that it has gynoecium because it has a stigma that is part of the gynoecium composition. We can also see that she has androecium, because she has an anther that is part of the composition of androecium. The flower also has a corolla and chalice, since the chalice is formed by the sepals and the corolla by the petals.

b. This is a perfect flower, as we can see that androecium and gynoecium are present in the same flower. Imperfect flowers are those with only androecium or gynoecium.

c. The flower has bilateral symmetry, which is common in all orchids. This type of symmetry allows the flower to only be divided into two equal parts. Radial symmetry, on the other hand, allows flowers to be divided into many equal parts.

What is the important organelle that eukaryotic cells have and prokayotic cells don't have?

Answers

There are several differences between the two, but the biggest distinction between them is that eukaryotic cells have a distinct nucleus containing the cell's genetic material, while prokaryotic cells don't have a nucleus and have free-floating genetic material instead.

What are the three
parts of the hair SHAFT?

Answers

Answer:

The hair shaft is comprised of three layers: the cuticle, cortex, and medulla.

Explanation:

Answer:

Hair shaft is consisted of three layers: cuticle, cortex and in certain cases medulla. Flat and square-shaped cuticle cells are adhered tightly to the cortex cells proximally.Explanation:

list the 3 main types of rocks and the process of how they are craeted

Answers

There are three main types of rocks: sedimentary, igneous, and metamorphic. Each of these rocks are formed by physical changes—such as melting, cooling, eroding, compacting, or deforming—that are part of the rock cycle. Sedimentary rocks are formed from pieces of other existing rock or organic material.
sedementarj metaporhic and ignojus are ur andwer

To survive, an organism must be able to maintain stable internal conditions in a changing environment. This process is called homeostasis. The Human Homeostasis Gizmo™ allows you to explore how the human body stays at a nearly constant temperature in different conditions. Notice the Air temp. and Body temp. thermometers representing the air temperature and body temperature.
1. What is the initial air temperature?
2. What is the initial body temperature?
3. Next to each factor listed below, write "increase," "decrease," or "same" based on how you expect that factor to affect body temperature.
A. Raising air temperature: __same____Sweating: __decrease___.
B. Lowering air temperature: ___increase___Shivering: __increase___.
C. Adding clothing: __increase_____Exercising: ___increase_____.

Answers

Answer:

1. Initial air temperature: 0 °C (32 °F)

2. Initial body temperature: 37 °C (99 °F)

3. The expected effect of different factors on body temperature are:

   A. Raising air temperature: Increase

   B. Lowering air temperature: Decrease

   C. Adding clothing: Increase

   D. Exercising: Increase

Explanation:

The question is incomplete, it is necessary to look for a software related to this homework.

Homeostasis is a property of organisms that consists in their capacity to maintain a stable internal condition by compensating for changes in their environment through the regulated exchange of matter and energy with the exterior. It is a a dynamic equilibrium controlled by a mechanism of feedback that constitute the self-regulating mechanisms. Examples of homeostasis are temperature regulation and the balance between acidity and alkalinity (pH)

The Human Homeostasis Gizmo™ is a software that allows you to explore  how the human body stays at a nearly constant  temperature in different conditions.  To use it, you have to set the air temperature at 0 °C (32 °F) and the body temperature at 37 °C (99 °F), and click Play. After a simulation of 1 hour, then click Pause and gather the information. Then, do the same cprocedure  to test the effect of other factors and Click Reset between each trial. To determine the effect of different factors on body temperature, you have to compare the  final body temperature to the final body temperature while  there is no specific movement or change.

Then:

1. Initial air temperature: 0 °C (32 °F)

2. Initial body temperature: 37 °C (99 °F)

3. The expected effect of different factors on body temperature are:

   A. Raising air temperature: Increase

   B. Lowering air temperature: Decrease

   C. Adding clothing: Increase

   D. Exercising: Increase

This is for a logical reason, anything you do to heat it or generate heat, will increase the temperature. And anything you do to generate cold, will lower the temperature.

The effect of different factors on body temperature according to Gizmo:

   A. Raising air temperature: Increase

   B. Lowering air temperature: Decrease

   C. Click Add to a sweatshirt, hat, pants, and  parka. Then, adding clothing:

   Maintained

   D. Set the Exercise level to 70%, then exercising: Increase

When the temperature increases, it is because you are able to record the results from the software and you see temperature goes from 99 °F to 100 °F. When the temperature decreases. it is because it goes from 99 °F to 97 °F. When it is maintained, there is no change.  

A scientist observes an object approximately 11 km in length with an elliptical orbit around the Sun. The object is made of dust, ice, and rocky materials and passes Earth every 75 years.

Answers

Answer:

Comet

Explanation:

Comet is made up of dust,ice and rocky materials and passes every 75 years


How can photosynthesis be related to cellular respiration?

Question 8 options:

These two processes have nothing in common.


They both have the same reactants (need the same things).


They both produce the same products.


What's produced (made) by one is needed (the reactants) for the other.

Answers

What’s produced by one is needed for the other

can you please help me ​

Answers

Answer: 27. arcuate    28. enzyme. 29. three regions

Explanation:

Складіть ланцюги живлення, починаючи з продуцентів і закінчуючи редуцентами:
а) сосна звичайна, жук-мертвоїд, хрущ (майський жук), бактерії гниття, шпак, плітка, неясить
сіра, мідянка;
б) рак, карась, павук-сріблянка, чапля сіра, ропуха озерна, щука, денні черви, хара ламка.

Answers

Answer:

i dont speak ur language sorry

Explanation:

In pea plants, flower color is controlled by a pair of alleles where purple (P) is dominant to white (p).

a. True
b. False

Answers

Answer:

True

Explanation:

An allele refer to two alternative forms of the same gene. An allele is a pair of genes found on a given chromosome. Usually, every allelic pair has one which is dominant and one which is recessive.

The dominant allele is expressed while the recessive allele is not expressed.

The colour of the pea plant is controlled by a pair of alleles wherein purple (P) is dominant to white (p).

The expressed colour of the pea plant(phenotype) is dependent on the arrangement of the genes(genotype) of the pea plant.

Providing an example from your day-to-day life, explain the functions of the nervous system.

Answers

The main functions of the nervous system are sensory input, information processing, and motor output.

HELP PLEASE
What behavior allows newborn animals to recognize and follow the first moving object they see?

Question 4 options:

conditioning


courting


mimicking


imprinting

Answers

Answer: imprinting

Explanation: i took the test

What are the key things to learn about a cell?

Answers

Answer:

1

Explanation:

all living organisms are made up of cells.

If a diploid number is 42, then the haploid number is…

Answers

Answer:

21

Explanation:

diploid number will be double the haploid number

Answer:

21

Explanation:

For the organism in this example the diploid is 42 making the haploid would be half of that or 21 chromosomes.

Happy learning! Brainliest if this answer helped!

Can someone help me ASAP!!!

Answers

Answer:

rrkufhjgfyrugf5td89yed/learningnice.com/ffiuvfk/answer

Explanation: u can find the answer there

HELP!!! I need the answer and reasoning!!! I’ll mark as brainliest!!!

Answers

Answer:

A. Destruction of terrestrial biota

Explanation:

all the other answers would decrease the carbon output. thickening of ocean sediments reinforces a carbon reservoir (which stores carbon), reduction in the use of fossil fuels would reduce the amount of carbon emissions, increase in marine biota = increase in carbon storage because marine biota is also a carbon reservoir.

Destruction of terrestrial biota would cause excess output because terrestrial biota is a carbon reservoir, and all of that carbon would be released into the carbon cycle.

HELP ASAP !!!!! I WILL GIVE POINTS. Complete the following sentence.
The animal _____
industry provides nearly 2 million American jobs.

Answers

Answer:

livestock

Explanation:

Answer: agriculture

Explanation:

List 3 properties of the
hair's cuticle

Answers

Answer:

Cuticle, Cortex, MedullaBUTProperties include: SmoothnessFlatnessMalleability

Explanation:

These are the main three properties of the hair's cuticle.

Answer:

smoothness

Explanation:

elasticity, softness

Which statement describes an autotroph?
.
A) an organism that breaks down and obtains energy stored in dead organic matter
B) an organism that captures energy from sunlight or chemicals and uses it to produce food
C) an organism that is completely reliant on consuming other organisms to obtain energy
D) an organism that meets its energy needs by feeding on organic matter or other organisms

Answers

Answer:

B

Explanation:

I believe it's b it makes sense-

bye hope it helps

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