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13 Cards in this Set
- Front
- Back
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What developmental structure is the origin of Glial Cells?
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Ectoderm
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What are the three major types of glial cells?
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Astrocytes, oligodendrocytes, and microglia
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What is the role of glial cells after injury?
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Their phenotypes are critically altered resulting in neuronal repair regulation. In CNS, astrocytes and oligodendrocytes prevent regeneration of neurons. In PNS, schwann cells facilitate it.
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What is the only myelin producing cell in the brain?
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Oligodendrocytes
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What is the role of Astrocytes in the synapses?
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Control concentrations of neurotransmitters and ions in extrasynaptic space. Also releace factors which promote synaptic formation and function, neuronal growth, oligodendrocyte maturation, and formation and control of BBB.
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What is the difference between Direct and Indirect immunofluorescence methods?
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Direct only uses a primary antibody (directly conjugated to fuorochrome) whereas indirect also utilizes a secondary. Indirect immunofluorescence is more sensitive.
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What is the difference between monoclonal and polyclonal primary sera?
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Monoclonal targets only a single antigen whereas polyclonal can target multiple epitopes. Polyclonal give greater signal.
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What immune system cells produce antibodies and what is the shape of the protein?
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B-cells
Y shaped |
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Describe the meaning of Donkey Anti-Mouse IGM in a secondary antibody
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The Anti-body was grown in a donkey, and is directed against Mouse, which must be the species in which the primary antibody originates. IG=Immunoglobulin and M=defined by the heavy chain used in the antibody.
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Why do we Fix cells/tissues?
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Fixation is critical so the cellular distribution of antigen is maintained in its original position, so cellular morphology is maintained and so that the antibody has access to all necessary compartments.
Over-fixation can cause changes in conformation or modification of antigen, rendering it unrecognizable by the antibody. |
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Why do we permeabilize cells/tissues?
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This step is based mostly on the location of the targeted antigen whether in the cytoplasm or in the nuclear envelope. Detergents such as Triton-x100 are used to dissolve lipids from the cell membrane making it permeable to antibodies.
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Why do we block cells/tissues?
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NAS and BSA are used to occupy sites for proteins which are not the intended targets of the antibody. Since the antibody will preferentially bind to its target antigens, the blocking serums will not overcome this affinity.
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What are the three steps following fixing/permeabilizing/blocking?
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Primary antibody incubation, secondary antibody incubation, and mounting.
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