Immunological Techniques Flashcards
What is process in raising antibodies?
1) mice are immunised with target proteins
2) B cells are harvested + fused with tumour cells to form a hybridoma
3) hybridoma that produces antibody against target protein is selected + cloned
4) antibodies secreted by cloned hybridoma are harvested + used in immunological techniques
What is blood typing?
Is an example of agglutination reaction used to determine blood type
How does blood typing work?
Blood sample is mixed with antibodies raised against A, B or RhD antigens
Sample is visually checked for agglutination (blood cells sticking together)
Agglutination indicates presence of antigens in blood sample
What are blood groups?
Group A
Group B
Group AB
Group O
What is flow cytometry?
Is technology used to analyse proteins on cells that are in suspension
What are applications of flow cytometry?
Diagnosis (CD4 T cell count in HIV) diagnosis of haematological malignancies
Research => cell sorting + identification + analysis of immune cells
What is immunohistochemistry (IHS)?
Is used to show distribution + localisation of antigens in tissue sections using antibody antigen interactions
What is application of IHS?
Is used in diagnosis of cancer
What does ELISA do?
Quantifies amount of protein /antibody in liquid sample such as sera or tissue culture supernatants
What are 4 different types of ELISA?
Direct
Indirect
Sandwich
Competitive
How does sandwich ELISA work?
Add capture antibody Add sample Add detection antibody Add substrate Determine optical density => spectrophotometer
What is western blotting?
Is technique used to detect proteins
What are 4 steps involved in western blotting?
Sample preparation
Electrophoresis
Transfer to membrane
Stain for protein of interest
What are application of western blotting?
Diagnostic use
Research => cell signalling proteins + mechanism of action for cancer drugs
