L12/L13 Flashcards
(110 cards)
ADH stimulates release of _____
ACTH
describe a sandwich ELISA
coat wells of plate with primary (capture) antibody.
antigen binds to primary antibody.
secondary (detection) antibody, which is attached to an enzyme, binds to the antigen. this forms a sandwich around the antigen.
a substrate is added which the enzyme converts into a colored compound.
the amount of product (color) is proportional to the amount of antigen present.
advantages of monoclonal and polyclonal antibodies
monoclonal - high specificity and consistency. unlimited supply after cell line is established.
polyclonal - broad antigen recognition. high sensitivity. inexpensive to produce.
explain radioimmunoassay (RIA)
Radioactive ligand and unlabeled ligand compete for same antibody. Competition is basis for quantitation.
what is immunoprecipitation?
binding of antibodies to the protein of interest and then isolating the antibody-bound complexes through selection for the antibody
advantages and disadvantages of RIA
advantages:
extremely sensitive, can process large numbers of samples, quantifiable, easily automated.
disadvantages:
can’t determine if hormone measured has biological activity, also must get rid of radioactive material.
explain direct ELISA
the unknown sample with the antigen of interest is adhered directly to the well, followed by detection with a primary antibody
explain indirect ELISA
the unknown sample with the antigen of interest is adhered directly to the well. a primary antibody binds to it. then a secondary antibody binds to the primary one for detection.
explain competitive ELISA
The sample containing the target antigen is added to the plate.
A labeled version of the antigen is also added, competing with the unlabeled antigen in the sample for binding to the antibody on the plate.
The more target analyte is present in the sample, the less of the labeled reference antigen will bind to the antibody.
An enzyme-linked secondary antibody is then added, which binds to the labeled reference antigen and allows for color detection
describe the patch clamp technique
an electrophysiological method that allows researchers to record and measure the flow of ions across biological membranes through ion channels, either at the whole-cell or single-channel level. It involves creating a high-resistance seal between a glass micropipette and a cell membrane to isolate and study the electrical activity of a small “patch” of the membrane
https://www.youtube.com/watch?v=mVbkSD5FHOw
key features of inbred mice
homozygous at all genetic loci, so genetic variation is a controlled variable. but immune systems are still different
knockout mouse
Technique that specifically removes
expression of a gene from both the
mouse germline and somatic tissues.
transgenic mice
Technique to produce mice carrying
multiple copies of a gene that will be
overexpressed in the genome
how is inducible transgenics useful?
Inducible transgenic mouse models allow for the activation of genes in specific cells and tissues at specific times by administering a drug or some other external agent
5 hormones secreted by kidneys
erythropoietin, calcitriol (active vitamin D), renin, thrombopoietin, klotho
where is erythropoietin secreted from and what is its function?
excreted from extraglomerular mesangial cells. stimulates erythrocyte production.
function of calcitriol
active form of vitamin D. increases absorption of calcium/phosphate from GI tract
where is renin secreted from and what is its function?
secreted from juxtaglomerular cells. activates the renin-angiotensin system by producing angiotensin I and ultimately aldosterone synthesis
function of thrombopoietin
stimulates megakaryocytes to produce platelets
where is klotho secreted from and what is its function?
secreted from distant tubules in kidney. calcium, phosphate, and aging.
_____ affects sodium reabsorption in the nephron.
Aldosterone
__________ promotes tubular reabsorption of calcium, phosphate excretion and the synthesis of 1,25-dihydrocholecalciferol (the active form of vitamin D).
Parathyroid hormone
_______ is made by the juxtaglomerular cells and catalyses the formation of angiotensin I and ultimately aldosterone synthesis.
Renin
V2 receptor location and function
found in principal cells of renal collecting duct. controls the reabsorption of water from the renal tubules into the bloodstream