U1-KA1 Lab Techniques Flashcards
KA1- Lab Techniques
What is a Hazard?
A hazard is a source of potential harm
What is a Risk?
A risk is the likelihood that a hazard would cause harm
Types of Hazards
- chemical
- physical
- biological
- radiation
- psychological
What is Accuracy?
The closeness of a measured value to the true value
What is Precision?
The closeness of repeated measurements to one another.
What is Reliability?
The overall consistency of a measure
Overall achieved consistency
Calibration Curve
A standard curve is a method used to determine the concentration of an unknown solution
State the Separation Techniques
- centrifugation
- chromatography (paper/thin layer/affinity)
- protein electrophoresis
Features of molecules that aid in separation
- density
- solubility
- size and shape
- charge
Centrifugation
A centrifuge is a machine which spins at a high speed to separate out components at different densities
Chromatography
Chromatography is an application to separate amino acids, proteins and other mixtures.
- paper chromatography
- thin layer chromatography (TLC)
- affinity chromatography
Gel Electrophoresis
Protein electrophoresis uses current flowing through a buffer to separate proteins
Electrophoresis - Native Gel
Separates molecules by shape, size and charge.
Doesn’t denature molecules
Electrophoresis - SDS-Page
Separates molecules by size alone.
Denatures molecules passing through
How are antibodies produced?
Produced as part of the immune response of a vertebrate. They are produced by B lymphocytes, and are Y-shaped globular proteins.
What is an Antibody?
An antibody is specific to an antigen. Each B lymphocyte produces a specific antibody. A specific antibody binds with a specific antigen to render it harmless.
Monoclonal Antibodies
Stocks of antibodies with the same specificity, known as monoclonal antibodies, are required for immunoassay techniques.
Immunoassay
A biological test to detect the presence of a certain molecule through the use of an antibody or antigen.
Monoclonal antibodies are used for …
- diagnose and detect diseases
- cancer treatment
- prevention of organ rejection
- pregnancy testing
ELISA technique
In this test, antibodies linked to reporter enzymes cause a colour change in the presence of a specific antigen.
Two other properties of “Reporters”
- Chemiluminescence
- Flourescence
Bright field microscopy
A (living or dead) sample is mounted on a slide. Samples are often stained before being viewed.
Allows you to look at whole organisms, parts of organisms or thin sections of dissected tissue.
Western Blotting
Technique used to identify the proteins in a sample. This is done using SDS-Page to separate proteins which are blotted onto a membrane then probed with fluorescent labelled monoclonal antibodies.
