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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

i wante to study medicine in university so how i should prepare my self
Lissa Reply
microorganism functions
NUHU Reply
how does the helicobacteri pylori affect the stomach walls?
Erick Reply
hi
Allan
Hlo
SUMIT
hi
Umar
hi
Umar
when the stomach is been affected with helicobacteria what are the preventive measure to be considered to ensure specific outcome?
Umar
yeah
Ayan
what is microbiology
vijay Reply
microbiology is the scientific study of microorganisms .microorganisms are those organisms that are too small to see with the naked eye ex-bacteria fungi
Yashkin
a branch of biological science concerned with organisms that can not be observed with a naked eye
Mooya
what are the types of granulocytes and explain
lord
polymorpho nuclear leukocyte, known as granulocyte are divide into three,1-polymorpho eosinophil 2-polymorpho basophil 3-polymorpho neutrophil
Musa
microbiology is scientific study of microorganism which can nt be seen by naked eye,for example bacteria,viruses, protozoa ad fungi.
Emma
microbiology is the scientific study of microorganisms
Atia
I want to know more about sample collection on the field
Ama Reply
blood collection and urinarysis
Lizzy
2143
Lizzy
yes
Lizzy
In the periodic table the number on the upper left hand side is what
Aurelia Reply
Hydrogen
Tob
am not talking about the elements
Aurelia
Is it the atomic number or the mass number
Aurelia
hologen
Usman
differences between acid fast and non acid fast bacilli
ANTHONY Reply
acid fast have cell wall that holds to carbol fuschin stain while non acid fast doesn't have. it readily releases out the primary stain the carbol fuschin.
LAFIA
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eklectc
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kennedy
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Olivia
for sure, this question is not related to the topic.
LAFIA
can someone explain the process of glycolysis and the electron transport chain? I'm so freakin lost. it loses carbons, gains hydroxyls, gains, loses Hydrogens....ugh it's like a foreign language to me! or direct me to a youTube video or something that will make this seem easier to concept?
eklectc
it's a loaded question, sorry!
eklectc
why is DNA a genetic material
Mcbeth Reply
DNA is genetic material because it contains chromosome contains the traits which includes characters and behavioral characteristics
chima
why is it difficulty to classfy protista
Tanaka
Good
Eddy
what is infection prevention
Muhammed Reply
good hygiene
Dhaqan
way of preventing disease causing germs
henry
maintenance of sterilization
Pooja
h
Faustina
describe the components of the epidemiology triangle
Muhammed Reply
Hai
Nantongo
hii
Md
where from you
Md
i am Indian
Md
you
Md
Hello friend
effiong
How are you people doing
effiong
أ‌) Host factor ب) pathogen ج) environment
Widad
Hello
Kofi
Hi
Widad
hey hi
kalai
The epidemiologic triangle is made up of three parts: agent, host and environment
Princess
please can a microbiologist will work at hospital
Usman
what are the fluids used in biochemistry Lab used to diagnose diseases
Jb Reply
fadumo qule a gemil3
fadumo Reply
Faadum mahamud disease micro biology
fadumo
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fadumo
history of microbiology
Balqees Reply
Penicillin is caused by what microorganism
Balqees
Penicillin is caused by what microorganism
Balqees
penicillium notatum
Pooja
M sorry I mean penicillin is caused by what Fungi
Balqees
penicillium fungi
Pooja
when I finish with my bsc in microbiology where wil I work
Usman
Why can we see ourselves in a mirror?
Usman
mention 5 characteristics of prokaryotic cell and also 5 characteristics of euryotic cell
Grace Reply
PROKARYOTES _ does not have nucleus _does not have membrane bound organels like eukaryotes -does not have endoplasmic reticulum _does not have a mitchochondrion _it have plasmid instead of chromosome EUKARYOTE S _have true nucleus _have all membrane bound organels _have mitochondria have
Pooja
continuation _have endoplasmic reticulum _have chromosomes does not have plasmid
Pooja
antigenisity define
kalai
explanation of spores
nahida Reply

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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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