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Chemistry Instrumentation
Exam 1
37
Chemistry
Graduate
07/08/2017

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Term
Ampholyte
Definition
a molecule that is positively or negatively charged on the basis of the pH of the solution.
Term
True or false: a protein is an ampholyte
Definition
true
Term
Electrophoresis
Definition
the migration of charged solutes or particles within a liquid medium under the influence of an electrical field
Term
Isoelectric Point
Definition
the pH at which the molecule has no net charge and will not migrate during electrophoresis
Term
Theory of Electrophoresis: what charges go where?
Definition
+ = cathode
- = anode
Term
Theory of Electrophoresis: Rate of Migration
Definition
1. net charge of the molecule
2. size/shape of molecule
3.electric field strength
4. properties of supporting medium
5. Temperature
Term
Wick flow
Definition
The result of electrophoretic process that generates heat, causing evaporation of solvent from the electropheretic support.
Term
Electrophoretic mobility
Definition
the rate of migration per unit field strength
Term
What is the Mobility equation?
Definition
Mobility = Q/6rπ
Term
EM:
1. ____________ related to size of the molecule,
2. _____________ related to Q (net charge)
Definition
1. inversely
2. directly
Term
What are the four components of an electrophoresis system?
Definition
1. Two buffer chambers or "buffer plates"
2. electrodes
3. Electrophoresis support
4. Wicks
5. Cover
Term
Electrophoretic Stain: Protein Serum
Definition
Amido Black, Coomasie Blue, Ponceaus
Term
Electrophoretic Stain: Isoenzyme Ormazan
Definition
Nitrotetrazolium Blue
Term
Electrophoretic Stain: Lipoprotein zones
Definition
Fat Red 7B, Oil Red O, Sudan Black B
Term
Electrophoretic Stain: DNA fragments
Definition
Ethydium Bromide
Term
Electrophoretic Stain: CSF proteins
Definition
Silver Nitrate
Term
Electrophoretic Techniques:

Slab Gel
Definition
separation of serum of CSF proteins and DNA/RNA fragments; horizontal or vertical
Term
Electrophoretic Techniques:

Disc
Definition
5 zones, 20+ fragments, "discontinuities"
Term
Electrophoretic Techniques:

Isoelectric focusing
Definition
separates amphoteric compounds; sharp protein zones
Term
Electrophoretic Techniques:

2D
Definition
Used to study families of proteins, searches for genetic or disease
Term
Electrophoretic Techniques:

Southern Blot
Definition
used to detect fragments of DNA; uses AGE; used to (1) identify a DNA sequence, (2) determine the presence, position, and number of copies of a genome, and (3) typing DNA
Term
Electrophoretic Techniques:

Northern Blot
Definition
RNA; RNA probe used for hybridization
Term
Electrophoretic Techniques:

Western Blot
Definition
Protein; used to (1) separate, (2) detect and (3) identify one or more proteins in a complex mixture
Term
Electrophoresis: purposes of a buffer
Definition
1. carry current
2. Establish the pH
3. Determine electrical charge of solute
Term
Electrophoresis:
What provides the matrix in which separation takes place?
Definition
The support medium
Term
Electrophoresis:
Separation is based on what?
Definition
differences in charge to mass
size of molecule
Term
Electrophoresis:
Agarose separates what analytes?
Definition
separation of
-Serum, urine, or CSF proteins
-hemoglobin variants
-Isoenzymes
-Lipoproteins
Term
Electrophoresis:
Advantages of Agarose
Definition
Lower affinity for proteins, native clarity for drying, free of ionizable groups, preventing it from endosmosis
Term
Endosmosis
Definition
Highly hydrated ions in the applied current of the system move toward the electrode of the opposite polarity which causes movement of the solvent
Term
Zone Electrophoresis
Definition
Produces zones of proteins
Term
Electrophoretic Techniques:

Microchip
Definition
alternative to capillari electrophoresis; DNA separations
Term
Go through the different Electrophoresis Clinical Significances
Definition
Extra Electrophoresis Packet
Term
a1 zone
Definition
Increases: inc. in a-1 AT
Decreases: Bence Jones protein binding (RARE)
Term
a2 zone
Definition
Haptoglobin, a2 macroglobin, HDL
Term
beta zone
Definition
Represents:
Transferrin
Beta-Lipoprotein
C3 Complement
Elevated: High LDL, Pregnancy, Elevated C3, Increase in IgA
Term
Gamma Zone
Definition
In Order: IgG IgA IgM IgD IgE
Single band : Monoclonal Gammopathy
Term
Monoclonal Gammopathy of Undetermined Significance (MGUS)
Definition
condition in which an abnormal protein is in the blood
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