|Quantitative RT-PCR [110,111,112]||- Very specific quantitative information on mucin expression at the mRNA level|
- Inexpensive and easily applicable to most samples.
|- Inability to detect increase in secretion.|
- Post-transcriptional modifications are also not detected.
|Northern-blot (RNA-blot) assay [110, 111]||- Alternative method for detection of RNA.|
- Allows for separation of RNA molecules by size.
- Provides information on number, length, and relative abundance of mRNAs expressed by a single gene
|- More laborious, time-consuming and not as sensitive as qRT-PCR.|
- Requires large amount of tissue/sample, and high purity and quality of non-degraded RNA, which can be difficult for the large RNA molecules of mucins.
|Luciferase reporter and Chromatin immunoprecipitation (ChIP) assay (promoter-binding) [111, 113]||- Luciferase reporter assay is commonly used to study gene expression at the transcriptional level.|
- ChIP allows for the specific study of molecular regulation and induction of mucin expression under various conditions.
|- Most applicable in cell cultures.|
- Does not give quantitative information on mucin expression or secretion.
|Using transgenic or knockout animals [89, 114, 115]||- Unique and valuable information on the overall function and/or effects of overexpression/depletion of each mucin throughout the lifespan of an animal model.|
- Can be used for determination and verification of mucin-regulation pathways.
|- Most often applied in rodents.|
- Often have to be used in tandem with other techniques to verify the effect.
- Depending on the model species, can be expensive, time-consuming.