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Analysis and handling of IHC common problems
本页内容
Problem
Reason
Suggest
The result is too dark
The concentration of the primary antibody is too high or the incubation time is too long
Reduce primary antibody concentration or incubation time
The result is too dark
The incubation temperature is too high
Incubate at room temperature or 4 °C
The result is too dark
The incubation time of the HRP-labeled secondary antibody is too long
Reduce time
Non-specific staining in IHC results
The paraffin section is not completely dewaxed
Extend deparaffinization time
Non-specific staining in IHC results
Insufficient rinsing during the procedure
Increase the number and duration of washes
Non-specific staining in IHC results
The tissue is rich in endogenous biotin and peroxidase
Block with relevant reagent
Non-specific staining in IHC results
Antigenic ectopia occurs
Non-specific staining in IHC results
Insufficient protein blocking
Increase the protein blocking time
Weak color development in IHC results
The concentration of the primary antibody is too low or the incubation time is too short
Increase the concentration of the primary antibody or extend the incubation time
Weak color development in IHC results
The reagent has expired
Confirm the shelf life of the reagent before the experiment
Weak color development in IHC results
Excessive rinsing during the procedure
Rinse appropriately
No staining in IHC results
Expression of non-target antigens in the tissue
No staining in IHC results
mismatch between the species of the primary antibody and the secondary antibody
Verify the species origin of the primary antibody before the experiment
No staining in IHC results
mismatch between the chromogen and the HRP system
Verify the color development system before the experiment
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