Biological Tools
We have developed many tools in recent years to study mucus and mucins. Among them, here are a few that might be useful for your research.
Antibodies
Anti mouse-Muc6 antibody (rabbit polyclonal). Directed against the peptide TSKIPVTQTTTHRVPSRCIT, which is N-terminal to the tandem repeats of Muc6 (amino acids 1,387–1,406, GenBank: AY184388.1). For WB and IHC, available upon request.
Serial sections showing strong labelling of the deep gland cells with the anti-Muc6 antibody (CP4), and Muc6 within the gel layers, while Muc5ac was synthesized in the superficial/foveolar epithelium in agreement with the human distribution.
This antibody has been used in the following studies:
♦ Histochem Cell Biol. 2010; doi: 10.1007/s00418-010-0688-8
♦ Histochem Cell Biol. 2019; doi: 10.1007/s00418-019-01785-5
Anti mouse-Muc5b antibody (rabbit polyclonal). Directed against the 19 aa peptide DVSSGLVCYNKDQGGTFQM found in CYS domains #2/3/5 of Muc5b. For IHC, available upon request.
← Immunofluorescence showing Muc5b (green) production in bronchi. This antibody has been used in the following studies:
♦ Histochem Cell Biol. 2011; doi: 10.1007/s00418-011-0872-5
♦ PLoS One 2012; doi: 10.1371/journal.pone.0046699
♦ Sci Rep. 2017; doi: 10.1038/srep44591
♦ Histochem Cell Biol. 2019; doi: 10.1007/s00418-019-01785-5
♦ Biol Open 2019; doi: 10.1242/bio.046359
♦ iScience 2022; doi: 10.1016/j.isci.2022.104526
Mini5B expression plasmid
Mini5B expression plasmid: Eucaryotic hybrid promoter + intron + Mini5B + Ires-Luc. The plasmid contains a Neo cassette for stable transfection. The mini5B is made of two MUC5B CYS domains flanking a full TR array of MUC5B with a functional N-glycosylation site at the C-term.
This construct with or without the Ires-Luc cassette has been used in the following studies:
♦ PLoS One 2012; doi: 10.1371/journal.pone.0046699
♦ Biochem Biophys Res Commun. 2018; doi: 10.1016/j.bbrc.2018.10.138
Transgenic mice
Tff3-rCYSx12-GFP
Tff3-rCYSx12-eGFP. Mouse Tg: Tff3 promoter driving the production/secretion of a meGFP-tagged CYSx12 (human MUC5B) recombinant molecule with Muc2 in the mouse intestine. With its large Tff3 (mItf) promoter, this transgenic mouse can be used as a Tff3 eGFP-reporter mouse. Available under a collaborative MTA. Official name: Tg(Tff3/MUC5B/GFP)222
Immunofluorescence showing rCYSx12-eGFP (green) secreted with Muc2 (red) in the colonic mucus gel layers. This transgenic mouse has been used in the following studies:
♦ Sci Rep. 2015; doi: 10.1038/srep09577
♦ Sci Rep. 2019; doi:10.1038/s41598-019-53547-x
♦ Gut Microbes 2020; doi: 10.1080/19490976.2020.1800897
♦ iScience 2022; doi:10.1016/j.isci.2022.104526
Tff3-rCYSx12. The variant without the eGFP tag (named Tg208 or K208) has been deposited and is available at EMMA (B6;D2-Tg(Tff3/MUC5B)208Jlcd/Cnbc). Homozygotes are viable. For details, see MGI/Jax.
Reporter Muc5b-meGFP and derived Muc5b knockout
Muc5btm2.1Jlcd ; Muc5btm2.2Jlcd
Mouse eGFP reporter for Muc5b obtained by homologous recombination (knockin). The full Muc5b glycoprotein is labeled with a monomeric eGFP tag. The last two exons of Muc5b are floxed, giving the opportunity to generate any conditional Muc5b knockout. Available under a collaborative MTA.
These transgenic mice have been used in the following studies:
♦ Sci Rep. 2017; doi: 10.1038/srep44591
♦ PLoS One 2017; doi: 10.1371/journal.pone.0174764
♦ Histochem Cell Biol. 2019; doi: 10.1007/s00418-019-01785-5
♦ iScience 2022; doi: 10.1016/j.isci.2022.104526
Muc4 knockout
Muc4 knockout. Mouse Muc4 knockout was obtained by gene targeting. The first exon was removed. This transgenic model has not been published. Available @ Infrafontier/EMMA. Stock EM:15255
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