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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 morphology
Lovely Reply
study of external features
Katta
i think it is about characteristic that we see on the organisms like the shape
Aisyah
morphology is something is it's form and structure.
Rose
yes it's a study deals with the form of living organisms, and with relationship between their structure s.
Rose
is the study of size and shape of microorganisms
Elinam
what is mutation?
mavis
mutations is a change in the base sequence of DNA
Elinam
what's gram negative and positive bacteria
Dagbanja Reply
is a process that classify bacteria structure
moshtak
how
Dagbanja
1)the crystal voilet dye stains all cells blue 2)the iodine solution two groups are blue 3)alcohol such as acetone or ethanol gram negative colorless while gram positive remains blue 4)the red dye safranin gram positive reamin blue while gram negative red.
moshtak
gram negative and Gram positive bateria aare grouped on the nasis of the thickness of their peptidoglygan gram negative bacteria hava thin layer of petidoglycan while Gram positive bacteria has thick petidoglycane layer
Samina
hence during gram stainig after additon of iodine on crystal voilet it forms crystalvoilet-iodine complex, when we add acetone, due to thin peptidoglyacan layer the Gram negative bateria easily leaves this complex and becomes colourless while the gram positive bateria remains blue...
Samina
after addition of counter stain such as saffranin, the Gram negative gets stained pink
Samina
why are there so many long words and is there simplified versions of these words?
Ethan Reply
What is the different between abiogensis and biogensis
RAPHERA Reply
Abiogenesis is the process by wich living organisms arises from a nonliving matter while Biogenis organisms arises from living matter
Nji
Abiogenesis deals with the law of spountaneous generation which talks about organisms arising from non-living organic matters while biogenesis deals with organisms arising from living organic matters
Ademola
What are the importance of mircobiology
RAPHERA
mechanisms of microbiology
Erneet Reply
What is immunizations
RAPHERA Reply
the action of making a person or animal immune to infection, typically by inoculation.
Pup
pls someone should help with ANTIMICROBIAL MNEMONICS
Shuaib Reply
Aminoglycosides: Charasteristics of Aminoglycosides Nymonics; (AMINO)  Active against Aerobic gram negative Organisms Mechanism Of resistance is by Modifying enzymes Inhibit protein synthesis by binding to 30s ribosomal subunit Nephrotoxic Ototoxic Members  Of Aminoglycosides: Nymonics: (ST
Najeebul
Please can some briefly describe the germ theory?
Michael Reply
hyy all
Shuaib
pls someone should help with ANTIMICROBIAL MNEMONICS
Shuaib
which drug can be used to treat gonococus?
Lambon Reply
Madication - Antibiotic " and Gonorrhoea can be treat with Antibiotic
Maryam
Penicillin -An antibiotic
Michael
Ciprofloxacin tablets also an antibiotic
Michael
Ofloxacin is also antibiotics
Ikenna
yes
Elli
sory no fot gonococus
Elli
oofer gang roblox memes
Ethan
what are eukaryotes
Kakande Reply
eukaryotes simply refers to a cell who does not have a definite nucleus.
Franca
Eukaryotic cell has a true nucleus
arfa
what is right answer ?
krishna
I can remember now truly that eukaryotes have true nucleus.I was referring to procaryotes.
Franca
Eukaryotes have got true nuclues
Ede
they have a nucleus, larger than prokaryotes
Alanna
The eukaryotes cell are found in advance organisms.
Franca
Eukaryotes are multicellular organism which has a membrane bounded structure mostly found in animals and some plant cells
KINGSFORD
bacteria growth curve
sani Reply
No bacteria's don't grow curvey. Curve is a shape for a bacteria not growth
KINGSFORD
What is the difference between eukaryotes and prokaryotes?
KINGSFORD Reply
What's a multicellular organism?
KINGSFORD
multicellular organisms is an organism composed of many cells which are to varying degree integrated and independent.
Ruth
Boiling of milk to kill germs is called?
Kasigwa
pasteurization
Aiyedun
pasteurization
Katta
kadha
Katta
pasteurization
Gomathi
what is microscopic
monish
Pasteurization
Hosy
what are Flora
Hosy
you mean like the normal flora?
Tenucharity
pasteurization has to do with killing of the harmful microbes present for safe consumption. Eg. pasteurized yoghurt beside the bacteria that are supposed to be present, there may be harmful ones that have been killed
Tenucharity
Can someone explain flora
KINGSFORD
what are the normal Flora on the ff anatomical sites? Skin, nose, ear,oral cavity, intestines and the vagina
Hosy
what is cell?
Anita Reply
the cell is the functional and structural unit of the body
Ruth
yes ur right
Kasigwa
who add me
Wali
hy
Katta
hy
Shivangi
basic structural, functional, and biological unit of all known living organisms. A cell is the smallest unit of life.
Haru
hi
Ruth
hi
Haru
coming Oct 2 I have a exam on microbiology
Haru
okay all the best
Ruth
hi
sachin
hi
Ruth
plz bro send question of microbiology
umesh
structural and functional unit of the life
monish
this is not the correct answer
Alanna Reply
why
Hayat
why not
Zaajid
what is the correct answer?
vannessa
where is question
Zaajid
ask you question
Zaajid
What are mitochondria
Hayat
what is mitochondria
Hayat
who is the father of microbiology
T-
not
Hayat
Antonie Van Luweenhoek
Ravikumar
Mitochondria is the powerhouse of the Cell. And is the cell organelle which helps cell in generation of energy by Cellular Respiration
Ravikumar
Mitochondria are threadlike structuresbounded by double membrane,and serves as the power house of the cell
Felicity
can I get notes on introduction to microbiology just started I have no idea please
rita
Mitochondria is a membrane bounded structure, it's sausage in shape and has inner and outer membrane. The inner membrane is folded at the surface called cristae which project to form matrix. They function as the power house for the cell.
KINGSFORD
Rita are you a medilab student?
KINGSFORD
the father of microbiology is Luis pasture
Able
Erythromycin--------(+++) Cefuroxine ------------(++++) Chloramphenicol --(++) Vancomycin ---------(++++) please what does the above result means. under urine culture and sensitivity?
Kwame
francesco redi was the first to challenge spontaneous generation not pasteur
Alanna
Rita are you a medilab student? no please am a public health student
rita
kwame Evans this result ll mean that the bacterial in the urine can be remedy / killed by this antibacterial (cefuroxime and the rest are antibiotics
rita
Rita Appiah thanks.
Kwame
contact backstage
Emmanuel
what is a gram negative bacteria
Dagbanja
what's agram positive bacteria
Dagbanja
what is a vacual
Star 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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