Abstract
It is increasingly important to understand a gene’s function at the protein level. Immunohistochemistry is used to identify new functions of phytohormones by finding their precise localization patterns. To facilitate protein immunolocalization, we developed a protocol that can significantly reduce background and give highly specific signals of antibody to traget proteins. The key improvement is to treat secondary antibodies by mixing with homogenized fresh and fixed plant tissue. Using this protocol, we have successfully carried out immunolocalization of proteins encoded by genesAP3 andHoMADS2, as well as plant hormones ABA and IAA.
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Abbreviations
- AP3:
-
apatala 3
- AP:
-
alkaline phosphatase
- BS:
-
blocking solution
- BSA:
-
bovine serum albumin
- EDC:
-
1-(3-dimethylaminoprapyl)-3-ethyl carbodiimide
- FAA:
-
formaldehydeacetic acid medium
- HWB:
-
high-salt washing buffer
- LWB:
-
low-salt washing buffer
- PBS:
-
phosphate-buffered saline
- PI:
-
pistallata
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Gong, HQ., Peng, YB., Zou, C. et al. A simple treatment to significantly increase signal specificity in immunohistochemistry. Plant Mol Biol Rep 24, 93–101 (2006). https://doi.org/10.1007/BF02914049
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DOI: https://doi.org/10.1007/BF02914049