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troubleshooting:
Immunofluorescence staining is a very sensitive method that requires a lot of experience and optimization
| reason | solution | ||||||||||||||||
| Improper or too long fixation, resulting in artifacts | Reduce fixation time or change fixatives | ||||||||||||||||
| Insufficient closure | Extend the incubation time, consider using a different blocking solution (we recommend using serum from the secondary antibody host) | ||||||||||||||||
| Primary antibody is not specific | Use primary antibodies proven for immunofluorescence in the model system of choice; if available, use knockdown/knockout samples as negative controls | ||||||||||||||||
| The primary/secondary antibody concentration is too high, the incubation time is too long, and the incubation temperature is too high | To optimize antibody concentration and incubation time/temperature, consult manufacturer's protocol | ||||||||||||||||
| Cross-reactivity of secondary antibodies | Use isotype controls for secondary antibodies to check for cross-reactivity | ||||||||||||||||
| not washed enough | Verify that all wash steps are performed correctly; extend wash steps if necessary | ||||||||||||||||
| Bleaching during imaging | Use secondary antibodies conjugated to fluorophores suitable for the microscopy technique of your choice | ||||||||||||||||
| Low signal strength, causing noise | Optimize the signal-to-noise ratio, e. ibidi μ-Slides and μ-Dishes with coverslip bottoms for inverted microscopy are available in a variety of geometries to suit any specific need for your immunocytochemistry The geometry of the ibidi channel slides is ideal for the precise exchange of small intermediate volumes, which is required during immunocytochemical staining In ibidi chamber slides, silicone gaskets with removable chambers are mounted on standard slides Low signal or no signal
Guangzhou Keshite Scientific Instrument Co. If you are interested please contact us Tel: 020-38102730 Service QQ: 501747125 Email: info@kosterscience. Address: Room D, Room 140, North Tower, Junhui Building, Tiyu West Road, Tianhe District, Guangzhou Pay attention to the WeChat public account to get the latest information in time
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