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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

okay. Go ahead and ask
Blessing Reply
Industrial microbiology mcq
mohamed
Okay. What's your question?
Blessing
life arises from living matter or live organism.
Swami Reply
yes
sildra
I think live matter arises from non living matter
sildra
I dont think so...can u explain with an example
Manya
living maters made by non living matters
sildra
non living matters like stones? rocks?
Manya
eno
sildra
no
sildra
then?
Manya
cells are made by C N O minerals etc
sildra
I mentioned these as non living maters
sildra
that's all
sildra
cells are made up of those things but they originate from living things..
Manya
ok
sildra
Ok..good chat:-)
Manya
where are you from
sildra
thanks
sildra
India...u?
Manya
Tamil nadu
sildra
I am from Maharashtra
Manya
what about your studies
sildra
completed bsc.. preparing for msc entrance...wbu?
Manya
same
sildra
are you microbiologist
sildra
yes i am
Manya
good
sildra
what s the scope for micro in ur state?
Manya
did you find your college to higher studies
sildra
have to give an entrance exam for every college here...so lets c
Manya
food industries, medical lab, vaccine industries ,etc
sildra
hoping for pune University...wbu?
Manya
great!
Manya
is that centeral University right
sildra
what is your namr
sildra
hello
Udhaya
Hi
Maruf
Family kindly help me with this question? 1) Shortlist the configurative measurements of the following human anatomical ranges of÷ - Blood ( haemeglobin) in both male and female - Haematocytes in both male and female - Hepatocytes in both male and female - Lymphocyte / T. Lymphocytes in both male
Gifted
My names are Gift Mwale and am a Zambian. Kindly help me with this research which goes like this... 1) Shortlist the configurative measurements of the following human anatomical ranges of ÷ - Blood ( haemeglobin) in both male and female - Hepatocytes in both male and female - Haematocytes in both
Gifted
please what is the full meaning for TCDS
UDEME
please can anybody talk about brain tumour and its cure.
BELLO Reply
enlargement of the thyroid gland resulting in over production of hormone.
Kamal Reply
What can u say on Thyroid Cancer?
Abdulkareem
Please, talk about the thyroid cancer.
BELLO
explain the Grave's disease
John Reply
what is cell
Avi Reply
is unit of life
Kamaluddeen
Ok
mohamed
who is an industrial microbiologist
Cynthia Reply
I want to know the biochemical composition of bacteria
Josh Reply
It contains peptidoglcon, DNA nd RNA
Asiya
bacteriophage disadvantage
Momina Reply
disease due to __________ abnormalities are termed primary immunodeficiencies
Tayee Reply
Some primary immunodeficiencies are due to a defect of a single cellular or humoral component of the immune system.
Prince
Examples of primary immunodeficiencies include: chronic granulomatous disease, X-linked agammaglobulinemia, selective IgA deficiency etc
Prince
thank you
Nana
explain microbial mutation
Emerald
what is mutation
Cynthia Reply
alteration in genetic makeup
UDEME
Hi, I'm new here. I'm Bello Abdul Hakeem from Nigeria.
BELLO Reply
welcome on board
Kamaluddeen
Thanks brother I'm an undergraduate. I hope to study for MBBS.
BELLO
pls guys help me out
Linda
what is limitation of plate
Linda
limitation of plate load test . there are some that should be considered while performing load test which are given below, this test is usually performed on relatively similar plate ,usually 1 or 2 square foot area the reason is that the plate of greater are the economically not feasible
yemi
iam new here my name shafiu umar from Nigeria
Shafiu
thanks shafiu
gambo
how to differentiate Gram positive from Gram negative bacteria?
Faheem
by the cell membrane
Tim
with gram staining which has 4 steps
Freedom
Mechanism of bacterial pathogenicity
rashida Reply
Toxicity
Ibrahim
write a short note on Algea
najaatu Reply
green minutes plants organisms that are produced in turf.
Prince
Algae is a kind of a photosynthetic organism, which is usually grown in the moist areas. These are usually the simple plants that grow near to the water bodies. It contains a kind of chlorophyll pigments that act as a primary coloring agent.
Avi
they are eukaryotic and most lived in fresh water. they are photosynthetic that's, they contain chlorophyll and store starch in discrete chloroplast.
Prince
are bacteria important to man
just Reply
yes, depending on the type of bacteria .eg the normal florals, the latic acid batteries etc are important to man
Lois
I mean latic acid bacterial
Lois

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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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