LAB CMB

Question 1

GIVE THE STEPS FOR WESTERN BLOT

Answer 1

TISSUE PREPARATION, GEL ELECTROPHORESIS TO SEPARATE PROTEINS, TRANSFER TO MEMBRANE, BLOCKING, DETECTION

Question 2

PROTEINS MOVED FROM WITHIN THE GEL TO A MEMBRANE MADE OF [BLANK]

Answer 2

NITROCELLULOSE OR POLYVINYLIDENE DIFLUORIDE (PVDF)

Question 3

PLACED ON TOP OF THE GEL, WITH STACK OF FILTER PAPERS PLACED ON TOP OF THAT.

Answer 3

MEMBRANE

Question 4

IS PLACED IN BUFFER SOLUTION WHICH MOVES UP THE PAPER THROUGH CAPILLARY ACTION, BRINGING PROTEINS UP WITH IT.

Answer 4

ENTIRE STACK

Question 5

[BLANK] IS EXPOSED ON A THIN SURFACE LAYER FOR DETECTION

Answer 5

PROTEIN

Question 6

DETECTION TAKES PLACE IN 2 STEP PROCESS

Answer 6

1.PRIMARY ANTIBODY IS ADDED AT AN APPROPRIATE DILUTION AND INCUBATED WITH THE MEMBRANE. 2. MEMBRANE IS RINSED TO REMOVE UNBOUND PRIMARY ANTIBODY. The SECOND ANTIBODY IS ADDED

Question 7

MEMBRANE IS “PROBED” FOR PROTEIN OF INTEREST WITH A MODIFIED ANTIBODY.

Answer 7

DETECTION

Question 8

ANTIBODY IS LINKED TO [BLANK]

Answer 8

REPORTER ENZYME

Question 9

AS THE MEMBRANE IS ABLE TO BIND PROTEIN, STEPS ARE TAKEN TO PREVENT INTERACTIONS BETWEEN THE MEMBRANE AND THE ANTIBODY.

Answer 9

BLOCKING

Question 10

AFTER EXCESS SECOND ANTIBODY IS WASHED OFF, A SUBSTRATE IS ADDED

Answer 10

DETECTION (CONT’D)

Question 11

AN [BLANK] PRIMARY ANTIBODY CAN ALSO BE USED WHICH CAN BE DETECTED DICTATED BY X-RAY FILM AND DOES NOT REQUIRE THE SECONDARY ANTIBODY.

Answer 11

ISOTOPE LABELED PRIMARY ANTIBODY

Question 12

APPLICATIONS ON WESTERN BLOT

Answer 12

HIV TEST THROUGH HUMAN SERUM SAMPLE, BOVINE SPONGIFORM ENCEPHALOPATHY, LYME DISEASE, HEPATITIS B

Question 13

DENATURATION BUFFER ON SDS PAGE

Answer 13

TRIS - HCl PH 6.8, SDS, B- MERCAPTOETHANOL, BROMOPHENOL BLUE, GLYCEROL, ddH2O

Question 14

HAVE HIGH AFFINITY FOR PROTEINS

Answer 14

MEMBRANES