2012考研专业课自测试题及答案教育研究方法.doc

TECHNICAL SERVICE GUIDE: WESTERN BLOTTING
Customer Information:
Catalog #: ABCG2(BXP-9):sc-58225 Lot #:G1610
A. Problem and Previous Experience
1. What is the specific problem you are experiencing?
I cannot get the ABCG2 protein.
2. What size bands were expected and what size bands were detected?
72kDa bands were was detected except GAPDHA protein.
3. Was the blot blank or was a dark background or non-specific bands seen?
Nothing was detected except GAPDHA protein.
4. Did this same vial of product work in the past? What were the results?
all think they are not steady.
5. Did other lots of this product work in the past? Which lots? What were the results?
. We all think they are not steady.
B. Samples and Controls
1. From what species (animal) was the sample and what type of sample was used (cell lysate,
tissue, purified protein, etc.)? What lysis buffer was used?
mouse
2. How much sample was loaded on the gel?
two
3. If the blocking peptide was used, was it used as a negative control?
Yes
4. What positive and/or negative controls were used, and what were the results?
GAPDH band was clear and vivid.
C. Blocking
1. What were the blocking conditions (blocking solution, how long, what temperature. etc.)?
With Western Blot,we use 5% milk to block for one hour .
D. Primary and Secondary Antibodies
1. At what dilut