BLOTTING Flashcards
____ are techniques for TRANSFERRING DNA, RNA and PROTEINS onto a ____ so they can be ____
Blots
Carrier
Separated
Blotting often follows the use of a ___.
gel electrophoresis
Which blot is associated with what?
Southern blot
Northern blot
Western blot
Eastern blot
Southern blot - DNA
Northern blot - RNA
Western blot - proteins (immunoblot)
Eastern blot - post-translational proteins
___ is an ANALYTICAL METHOD that involves the IMMOBILIZATION of ____ on ____ before detection using ANTIBODIES
Protein blotting
proteins
membranes
The ___ technique is a PREREQUISITE for WESTERN BLOTTING
SDS PAGE
____ (____) is an ELECTROPHORESIS METHOD that allows PROTEIN SEPARATION by MASS
SDS-Page
Polyacrylamide Gel Electrophoresis
___ is a NEGATIVELY CHARGED ___ used to DENATURE and LINEARIZE proteins
Sodium dodecylsulfate (SDS)
detergent
True or False: Big proteins end up at the bottom of the running gel
FALSE. Big proteins do not travel as fast to the positive electrode
____ used to FORM a GEL that provides a ___ of ___ through which MOLECULES MIGRATE at DIFFERENT RATES
Polyacrylamide
matrix
pores
Ways to visualize protein bands (2)
Visualize the band under UV
Visualize by staining
Visualization of protein bands:
Most common stain
Most sensitive test
Coomassie Blue
Silver stain
In order to make PROTEINS accessible to ANTIBODY DETECTION, they are transferred onto a MEMBRANE made of ___ or ___
nitrocellulose
polyvinylidene difluoride (PVDF)
Types of transfer for proteins (2)
Diffusion transfer
Electro transfer
SDS page short guide (4)
SDS-PAGE
TRANSFER to a MEMBRANE
BLOCKING and REACTION with ANTIBODIES
DETECTION of BANDS
