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Rheum Boards Studying > Lab techniques > Flashcards

Lab techniques Flashcards

1
Q

Direct Elisa

A
  • Sample containing antigen is coated onto the plate well and the labeled antibody of known specificity is added
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2
Q

Indirect Elisa

A
  • Most common
  • Samples added to wells coated with a known antigen
  • Antibodies in the sample that bind to the antigen are detected using enzyme labeled anti human IgG
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3
Q

Sandwich (capture) Elisa

A
  • Plate wells are coated with a capture antibody targeting a specific antigen
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4
Q

Inhibition Elisa

A
  • Utilizes competition for labeled antibody between sample and well wall antigen
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5
Q

Advantages to Elisa

A
  • Rapid
  • Automatic
  • Multiple samples
  • Objective
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6
Q

Disadvantages to Elisa

A
  • Less specific than other assays
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7
Q

Immunoprecipitation

A
  • Preciptating a protein antigen out of solution using an antibody that specifically binds to that particular protein
  • Can be used to isolate and concentrate a particular protein from a sample containing thousands of proteins
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8
Q

Western blotting

A
  • Protein antigens are separated based on molecular weight by electrophoresis
  • The separated antigen is blotted onto a nitrocellulose membrane that is then intubated with patient or control serum
  • Bound antibody is identified by use of labeled anti human IgG
  • Antibody present in the patient serum is identified by comparison with control serum results
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9
Q

Advantages to WB

A
  • Does not require purified antigen for testing

- Allows for determination of fine specificities of antibody responses to multi subunit antigens

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10
Q

Disadvantages to WB

A
  • Time consuming
  • Expensive
  • Requires denaturing proteins prior to gel ELectrophoresis
    and will therefore not detect antibodies against conformational isotypes
  • Poor sensitivity to Ro and SCL-70
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11
Q

Southern blotting

A
  • Like WB but DNA instead of protein
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12
Q

Indirect IF

A

Tag is used to identify antibody on a substrate

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13
Q

Direct IF

A

Tag is used to identify antigen on a substrate

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14
Q

Disadvantages to IF

A
  • time consuming
  • Labor intensive
  • Requires skilled operators
  • Subjective
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15
Q

DNA microarrays

A
  • Collection of microscopic DNA spots attached to a solid surface
  • Hybridizes complimentart DNA or RNA
  • Can be used to look for mutations of snps and gene expression (RNA)
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16
Q

Transgenic mice

A
  • Pronuclear injection into a single cells of the mouse embryo containing desired DNA sequence (gene) which randomly integrates into the mouse genome
  • Used to insert new genetic information or over express endogenous genes
17
Q

Gene knockout mice

A
  • Targets specific gene by inserting a foreign sequence that renders the resultant protein non functional, knocking out the gene
18
Q

Most commonly studied mouse model of RA

A
  • Autoimmune arthritis induced by immunization with emulsion of complete Freund’s adjuvant AND type II collagen
    (as opposed to adjuvant arthritis which is injection with M TB and is used more for reactive arthritis)
19
Q

New Zealand Mixed Mice

A
  • Oldest model of lupus
  • Developed severe lupus like phenotype including LAD, splenomegaly, ANA, dsDNA ad immune complex mediated GN leading to kidney failure and death
  • Disease is strongly biased in favor of females
20
Q

MRL/lpr mice

A
  • Develop SLE like phenotype characterized by LAD due to accumulation of double negative CD4-/CD8- and B220+ T cells
  • Accelerated mortality rate and both males and females are significantly affected
  • High concentrations of circulating immunoglobulins including ANA, ssDNA, dsDNA, SM, RF, resulting in large amounts of immune complexes
  • Mutation is in Fas receptors leading to a defect in apoptosis
21
Q

BXSB mice

A
  • Severe with early onset in males (due to Yaa)

- GN

22
Q

ALeutian disease in mink

A
  • Highly contagious parvovirus
  • Immune complex disease that resembles SLE including GN
  • Have positive DS dna antibodies
23
Q

g/d mouse

A

Similar to MRl/Lpr with defect in fas ligand and the process of apoptosis

24
Q

Tight mouse (tsk)

A
  • Fibrillin 1 gene mutation

- Causes autosomal dominant scleroderma like disease with skin and in some cases visceral fibrosis

25
Q

Model of myositis

A
  • Murine polymyositis like disease is associated with infection with viruses such as coxsackie
26
Q

Model of spondyloarthripthy

A
  • B27 transgenic rat

Rheum Boards Studying (34 decks)

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