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參數(shù)資料
| 型號： | A701 |
| 廠商： | Electronic Theatre Controls, Inc. |
| 英文描述： | A701: Test slide, various cancers plus corresponding normal |
| 中文描述： | A701：測試幻燈片，各種癌癥加上相應的正常 |
| 文件頁數(shù)： | 10/10頁 |
| 文件大?。?/td> | 217K |
| 代理商： | A701 |
IHC Conditioning
Negative controls
without a primary antibody, without a secondary antibody, or without detecting reagents.
Why does my negative control show strong
signal
The signal is due to non-specific cross-reactivity of
detection reagents.
Dilute the secondary Ab.
Change species of secondary Ab.
Detection reagent only is omitted. Intrinsic tissue
enzyme activity is interfering with the reaction. Treat
tissue with reaction solution.
Possible cause of signal
Trouble shooting
The secondary Ab is binding non-specifically to the
tissue. Add 0.1% tissue-specific serum to the
secondary Ab.
Primary Ab only is omitted.
The detecting reagents are binding non-specifically to
the tissue. Block tissue with detection reagent.
The secondary antibody only is omitted.
Positive controls
A positive antibody with the test tissue, or the test antibody with a positive tissue.
Using an antibody known to react with the test tissue, or using the test antibody with cells known to contain
the antigen. Fixatives may have reduced access of the antibody to the antigen. Perform microwave target
retrieval procedure or protease digestion.
Why does my positive control have no
signal
In most cases, this is because the antibody is not
optimized or the tissue is not adequately treated.
The enzyme/substrate system is defective or
incompatible.
This can be confirmed by performing by dot blot test.
Possible cause of no signal
Trouble shooting
Ab may be too dilute.
Titrate Ab to determine the optimum dilution that gives
the best signal-to-noise ratio.
Secondary Ab does not recognize the primary
Ab.
Ensure secondary Ab is directed against the species of
primary Ab, e.g. anti-mouse secondary Ab for mouse
primary Ab
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