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a) A graph showing a large peak at alb and smaller peaks at alpha1, alpha2, beta, and gamma.  B) A photograph showing various white lines on a blue background. The top line is labeled anti-human whole serum, the next is anti-IgG, the next is anti-IgA, the next is anti-K and the next is anti-lambda. Each of these is near a band from both the normal and patient urine samples. Black arrows point to the patient’s sample next to anti-IgG and anti-k. A white arrow points to the band from the patient near anti-K.
(a) This graph shows normal measurements of serum proteins. (b) This photograph shows an immunoelectrophoresis of urine. After electrophoresis, antisera were added to the troughs and the precipitin arcs formed, illustrating the distribution of specific proteins. The skewed arcs (arrows) help to diagnose multiple myeloma. (credit a, b: modification of work by Izawa S, Akimoto T, Ikeuchi H, Kusano E, Nagata D)

Protein electrophoresis and the characterization of immunoglobulin structure

The advent of electrophoresis ultimately led to researching and understanding the structure of antibodies. When Swedish biochemist Arne Tiselius (1902–1971) published the first protein electrophoresis results in 1937, Tiselius, Arne, “Electrophoresis of Serum Globulin: Electrophoretic Analysis of Normal and Immune Sera,” Biochemical Journal 31, no. 9 (1937): 1464. he could identify the protein albumin (the smallest and most abundant serum protein) by the sharp band it produced in the gel. The other serum proteins could not be resolved in a simple protein electrophoresis, so he named the three broad bands, with many proteins in each band, alpha, beta, and gamma globulins. Two years later, American immunologist Elvin Kabat (1914–2000) traveled to Sweden to work with Tiselius using this new technique and showed that antibodies migrated as gamma globulins. Tiselius, Arne and Elvin A. Kabat. “An Electrophoretic Study of Immune Sera and Purified Antibody Preparations,” The Journal of Experimental Medicine 69, no. 1 (1939): 119-31. With this new understanding in hand, researchers soon learned that multiple myeloma, because it is a cancer of antibody-secreting cells, could be tentatively diagnosed by the presence of a large M spike in the gamma-globulin region by protein electrophoresis. Prior to this discovery, studies on immunoglobulin structure had been minimal, because of the difficulty of obtaining pure samples to study. Sera from multiple myeloma patients proved to be an excellent source of highly enriched monoclonal immunoglobulin, providing the raw material for studies over the next 20-plus years that resulted in the elucidation of the structure of immunoglobulin.

Electrophoresis showing 4 healthy patterns and 3 examples of those with myeloma. All 7 lanes contain multiple blue bands with a thick band labeled albumin. The myeloma samples have a dark band labeled gamma-globulin, while the healthy samples do not have a band here. A graph showing the total gamma-globulin amount (g/dl) in each of these samples. The healthy samples have approximately 0.1 while the myeloma samples range from 0.8 to 1.
Electrophoresis patterns of myeloma (right) and normal sera (left). The proteins have been stained; when the density of each band is quantified by densitometry, the data produce the bar graph on the right. Both gels show the expected dense band of albumin at the bottom and an abnormal spike in the gamma-globulin region. (credit: modification of work by Soodgupta D, Hurchla MA, Jiang M, Zheleznyak A, Weilbaecher KN, Anderson CJ, Tomasson MH, Shokeen M)
  • In general, what does an immunoelectrophoresis assay accomplish?

Immunoblot assay: the western blot

After performing protein gel electrophoresis, specific proteins can be identified in the gel using antibodies. This technique is known as the western blot . Following separation of proteins by PAGE, the protein antigens in the gel are transferred to and immobilized on a nitrocellulose membrane. This membrane can then be exposed to a primary antibody produced to specifically bind to the protein of interest. A second antibody equipped with a molecular beacon will then bind to the first. These secondary antibodies are coupled to another molecule such as an enzyme or a fluorophore (a molecule that fluoresces when excited by light). When using antibodies coupled to enzymes, a chromogenic substrate for the enzyme is added. This substrate is usually colorless but will develop color in the presence of the antibody. The fluorescence or substrate coloring identifies the location of the specific protein in the membrane to which the antibodies are bound ( [link] ).

Questions & Answers

what is prokaryotes
PRALHAD Reply
are micro-organisms that have no nucleus e.g bacteria
Foster
and they do not possess membrane bound organelles e.g mitochondria
Bharat
yes
Foster
they are single cellular organisms,
Rajasekhar
all the organelles are embedded in cytoplasm of the cell
Rajasekhar
They don't possess true nucleus
PREMI
they're unicellular organisms
Kamaluddeen
they are classified as bacteria and. archaea. bacteria have a cell membrane conposed of peptidoglycan and archaea is composed of psuedomu
Tara
pseudomurein
Tara
organelles are non membraneous as the nucleus
Tara
What is the biosynthesis of a bacteria cell
Adesina Reply
biosynthesis is multiple step enzymes catalyze process which occur in bacterial cell
Jayeeta
biological way of manufacturing food and utilizes it for growth, multiplication and development
Kamaluddeen
prokaryot
PRALHAD
what is microbiology
Kiirya Reply
which organism can not be seen necked eyes is knowm as microbiology
meet
microorganisms
Jayeeta
micro can be define as d smallest living organisms dat can not be see wit naked eyes only with the aid of a microscope 🔬
muhammed
bio means life logy means study
muhammed
biology can b define as the systematic study of living things
muhammed
microbiology can be define as the study of tiny organisms, that's according to my understanding
muhammed
Microbiology is the study of living organisms which cannot be seen by the naked eye but only with the aid of a microscope. These microorganisms can be classified as bacteria, fungi, micro-algae, protozoa and virus.
Tatz
microbes that can't seen in naked eyes. upto 100micrometer we can see in naked eyes but 1 microns or nanometers are invisible in naked eye.
Jayeeta
what is a cell
Oteri Reply
cell can be define as smallest indivisible organism in the body
muhammed
cell is the smallest unit which makes up the whole body
Yue
cell is smallest unit and there are two types prokaryotic cell and eukaryote cell. bacteria a prokaryotic cell which works independently.
Jayeeta
nice forum
muhammed
Which parasite competes with the host for vitamin B12 to cause pernicious anaemia
Abubakar
Diphyllobothrium latum for vit B12
Jayeeta
it is also fish tapeworm
Jayeeta
Diphyllobothrium sp feed vitB 12
Jayeeta
recent trends in microbiology of biofertilizer
Ykomalipriya Reply
using nano technology
Jayeeta
what is transposors
EMMANUEL Reply
Junk DNA, jumping gene.
Pradeep
transposon elements are jumping gene as DNA sequence move from one location to anothet
Jayeeta
Yes
Pradeep
what is microbiology
Usama Reply
Microbiology is the study of organisms, most of which are too small to be seen with the naked eye, including bacteria, algae, protozoa, and fungi, as well as acellular agents, such as viruses and prions.
digvijaya
what is gram +ve and gram - ve bacteria?
Sharma
gm- contain lipid layer so it take counter stain during gram staining and pink or red in color but gm + does have thick LPS layer.
Jayeeta
what year microbiology started
Kartik
Robert Koch was the first to detect microorganisms
Jayeeta
what appears microorganisms
Kartik
clear the questions please
Jayeeta
was it robert koch or robert hooke?
digvijaya
microbiology word came from microns. study microbes that not recognize in naked eyes.
Jayeeta
what are the branch of microbiology
Oyemade Reply
We can basically classify into pure and applied microbiology
Ozzie
what is arabinose operon and it's stucture
Pooja Reply
The L-arabinose operon, also called the ara or araBAD operon, is an operon required for the breakdown of the five-carbon sugar, L-arabinose, in Escherichia coli
Tharra
kya hum operon ko divide kar skate hain
Kartik
reactions if viruses to physical and chemical agents?
VICTOR Reply
All viruses contain the following two components: 1) a nucleic acid genome and 2) a protein capsid that covers the genome. Together this is called the nucleocapsid. In addition, many animal viruses contain a 3) lipid envelope. The entire intact virus is called the virion.
Asali
assume you have performed a gram stain on a sample of pus from a patients urethra. you see red, nucleated cells and purple rods. what can you conclude
Petty Reply
bacteria
ABUBAKAR
gram negative bacteria hence gonococcal infection
Kiirya
Study of micro organisation which we cannot see with naked eye...
shyam Reply
Despite their bad reputation, microbes are mostly beneficial or have a neutral effect on our lives. Microbiology is the scientific study of these microorganisms. Microorganisms are those organisms that are too small to see with the naked eye and include things like bacteria, fungi, and viruses.
Asali
yes
Dhirender
not 'things'...'organisms' like bacterias, viruses and fungi
Sinmi
simi, it's not bacterias but bacteria which is the plural form of the word while bacterium is the singular of it
Richard
bacterium is obvio in a singular form but plural of bacterium is bacterias
Sinmi
Richard is right it's bacterium which is the singular form and plural is bacteria. *Bacterias* doesn't exist.
Aakriti
that is microbiology which is deals with microbes only their structure
Kiirya
what is microbiology?
Sandeep Reply
what is microbiology
Glory
is the study of small microorganisms that can't be viewed by our naked eyes under a microscope
Nabukwasi
it's the study of microbes their classification, diversity, their cellular structure and processes, their applications and interaction with other organisms.
Chaitanya
what is EMP pathway
rajesh
which organism is implicated in meningoencephalitis
Peter Reply
Herpes virus
Carlos
Yeah
Laxmi
which mold is called dairy mold?
Gopinath
mushroomm
Top
Geotrichum candidum
Gopinath
hi
bio
Hlw
Laxmi
which infectious agent is the smallest
Sinmi
spore or virus
Anushree
prion ....it is a single protein in a misfolded form..there is also a theory based on it
Sinmi
yup...m sry
Anushree
yes it can be turned as a virus
Sinmi
which gram positive bacteria is responsible for food poisoning?
Sinmi
prion is not a living organism yet it is infectoois why
Jeevan
prions are virus like organism. and it is a misfolded protein that may cause disease to us.
Tharra
Clostridium perfringens, B.cereus
Sneha
Who is the father of the biology
Dhirender
I have one more question. . that who is the father of botany and medicine? ? can any one tell
Dhirender
define polar bond in simple way
Rashmi Reply
when there is formation of positive and negative poles i.e. one atom is +ve and other is -ve which further results in the dipole moment. Such bond is know as polar bond. It is a convalent bond only.
Aakriti

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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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