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Molecular analysis of proteins

In many cases it may not be desirable or possible to study DNA or RNA directly. Proteins can provide species-specific information for identification as well as important information about how and whether a cell or tissue is responding to the presence of a pathogenic microorganism. Various proteins require different methods for isolation and characterization.

Polyacrylamide gel electrophoresis

A variation of gel electrophoresis, called polyacrylamide gel electrophoresis (PAGE) , is commonly used for separating proteins. In PAGE , the gel matrix is finer and composed of polyacrylamide instead of agarose. Additionally, PAGE is typically performed using a vertical gel apparatus ( [link] ). Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins based on their net charges. Alternatively, proteins can be denatured and coated with a negatively charged detergent called sodium dodecyl sulfate (SDS) , masking the native charges and allowing separation based on size only. PAGE can be further modified to separate proteins based on two characteristics, such as their charges at various pHs as well as their size, through the use of two-dimensional PAGE . In any of these cases, following electrophoresis, proteins are visualized through staining, commonly with either Coomassie blue or a silver stain.

a) A diagram showing a globular protein with positive and negative charges undergoing SDS treatment. SDS denatures the protein (producing a linear product) and makes them uniformly negative in charge. B) The protein samples are then placed into the wells of an SDS_PAGE gel. One well is loaded with a molecular weight standard. The gel is then exposed to a power source that results in the top of the gel (near the wells) becoming negative charged and the bottom becoming positively charged. Proteins migrate through the gel from the negative to the positive sides. Small proteins travel through the gel faster than large proteins. The molecular weight standard includes fragments of known size and is used to estimate the size of sample proteins. In this example the standard has sizes of 216, 132, 78, 32 and 7. The other lanes have bands of various sizes. C) A photograph of an SDS-PAGE gel. Purple bands on a clear background.
(a) SDS is a detergent that denatures proteins and masks their native charges, making them uniformly negatively charged. (b) The process of SDS-PAGE is illustrated in these steps. (c) A photograph of an SDS-PAGE gel shows Coomassie stained bands where proteins of different size have migrated along the gel in response to the applied voltage. A size standard lane is visible on the right side of the gel. (credit b: modification of work by “GeneEd”/YouTube)
  • On what basis are proteins separated in SDS-PAGE?

Part 3

When Kayla described her symptoms, her physician at first suspected bacterial meningitis , which is consistent with her headaches and stiff neck. However, she soon ruled this out as a possibility because meningitis typically progresses more quickly than what Kayla was experiencing. Many of her symptoms still paralleled those of amyotrophic lateral sclerosis (ALS) and systemic lupus erythematosus (SLE) , and the physician also considered Lyme disease a possibility given how much time Kayla spends in the woods. Kayla did not recall any recent tick bites (the typical means by which Lyme disease is transmitted) and she did not have the typical bull’s-eye rash associated with Lyme disease ( [link] ). However, 20–30% of patients with Lyme disease never develop this rash, so the physician did not want to rule it out.

Kayla’s doctor ordered an MRI of her brain, a complete blood count to test for anemia, blood tests assessing liver and kidney function, and additional tests to confirm or rule out SLE or Lyme disease. Her test results were inconsistent with both SLE and ALS, and the result of the test looking for Lyme disease antibodies was “equivocal,” meaning inconclusive. Having ruled out ALS and SLE, Kayla’s doctor decided to run additional tests for Lyme disease.

  • Why would Kayla’s doctor still suspect Lyme disease even if the test results did not detect Lyme antibodies in the blood?
  • What type of molecular test might be used for the detection of blood antibodies to Lyme disease?
A photo of a bulls-eye rash; a red spot in the center and a red ring around that.
A bulls-eye rash is one of the common symptoms of Lyme diseases, but up to 30% of infected individuals never develop a rash. (credit: Centers for Disease Control and Prevention)

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Questions & Answers

how does the planets on our solar system orbit
cheten Reply
how many Messier objects are there in space
satish Reply
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Richard Reply
what are astronomy
Issan Reply
Astronomy (from Ancient Greek ἀστρονομία (astronomía) 'science that studies the laws of the stars') is a natural science that studies celestial objects and phenomena. It uses mathematics, physics, and chemistry in order to explain their origin and evolution.
Rafael
vjuvu
Elgoog
what is big bang theory?
Rosemary
what type of activity astronomer do?
Rosemary
No
Richard
the big bang theory is a theory which states that all matter was compressed together in one place the matter got so unstable it exploded releasing All its contents in the form of hydrogen
Roaul
I want to be an astronomer. That's my dream
Astrit
Who named the the whole galaxy?
Shola Reply
solar Univers
GPOWER
what is space
Richard
what is the dark matter
Richard
what are the factors upon which the atmosphere is stratified
Nicholas Reply
is the big bang the sun
Folakemi Reply
no
Sokak
bigbang is the beginning of the universe
Sokak
but thats just a theory
Sokak
nothing will happen, don't worry brother.
Vansh
what does comet means
GANGAIN Reply
these are Rocky substances between mars and jupiter
GANGAIN
Comets are cosmic snowballs of frozen gases , rock and dust that orbit the sun. They are mostly found between the orbits of Venus and Mercury.
Aarya
hllo
John
hi
John
qt rrt
John
r u there
John
hey can anyone guide me abt international astronomy olympiad
sahil
how can we learn right and true ?
Govinda Reply
why the moon is always appear in an elliptical shape
Gatjuol Reply
Because when astroid hit the Earth then a piece of elliptical shape of the earth was separated which is now called moon.
Hemen
what's see level?
lidiya Reply
Did you mean eye sight or sea level
Minal
oh sorry it's sea level
lidiya
according to the theory of astronomers why the moon is always appear in an elliptical orbit?
Gatjuol
hi !!! I am new in astronomy.... I have so many questions in mind .... all of scientists of the word they just give opinion only. but they never think true or false ... i respect all of them... I believes whole universe depending on true ...থিউরি
Govinda
hello
Jackson
hi
Elyana
we're all stars and galaxies a part of sun. how can science prove thx with respect old ancient times picture or books..or anything with respect to present time .but we r a part of that universe
w astronomy and cosmology!
Michele
another theory of universe except big ban
Albash Reply
how was universe born
Asmit Reply
there many theory to born universe but what is the reality of big bang theory to born universe
Asmit
what is the exact value of π?
Nagalakshmi
by big bang
universal
there are many theories regarding this it's on you believe any theory that you think is true ex. eternal inflation theory, oscillation model theory, multiple universe theory the big bang theory etc.
Aarya
I think after Big Bang!
Michele
from where on earth could u observe all the stars during the during the course of an year
Karuna Reply
I think it couldn't possible on earth
Nagalakshmi
in this time i don't Know
Michele
is that so. the question was in the end of this chapter
Karuna
in theory, you could see them all from the equator (though over the course of a year, not at pne time). stars are measured in "declination", which is how far N or S of the equator (90* to -90*). Polaris is the North star, and is ALMOST 90* (+89*). So it would just barely creep over the horizon.
Christopher
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Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
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