Immunofluorescent staining of ISH-stained tissues and paraffin sections was performed as described previously. Primary antibodies were diluted as follows: 1:200 rabbit anti-CDH1 (3195, Cell Signaling Technology, Beverly, MA), 1:250 mouse anti-CDH1 (610181, BD Transduction Laboratories, San Jose, CA), 1:100 rabbit anti-phosphoSMAD1/5/8 (9511, Cell Signaling Technology, Danvers, MA). Secondary antibodies were diluted as follows: 1:250 Dylight 549-conjugated goat anti-rabbit IgG (111-507-003, Jackson ImmunoResearch, West Grove, PA), and 1:250 Dylight 488-conjugated goat anti-mouse IgG (115-487-003, Jackson ImmunoResearch). Immunofluorescently labeled tissues were counterstained with 4’,6-diamidino-2-phenylindole, dilactate (DAPI), and mounted in anti-fade medium (phosphate buffered saline containing 80% glycerol and 0.2% n-propyl gallate). Whole mount immunohistochemistry was performed as described previously. Primary antibody was diluted 1:750 rabbit anti-CDH1 and secondary antibody was diluted 1:500 biotin conjugated goat anti-rabbit IgG (BA-1000, Vector, Burlingame, CA).
Dylight 549 conjugated goat anti rabbit igg
Manufactured by Jackson ImmunoResearch
Sourced in Cameroon
DyLight 549-conjugated goat anti-rabbit IgG is a secondary antibody used for detection and visualization in immunoassays. It is produced by conjugating DyLight 549 dye to purified goat anti-rabbit IgG antibodies. DyLight 549 is a bright, photostable fluorescent dye that can be excited at 561 nm and detected in the orange-red region of the visible spectrum.
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Lab products found in correlation
Rabbit anti cdh1, by Cell Signaling Technology (2 mentions)
Mouse anti cdh1, by BD (2 mentions)
Biotin conjugated goat anti rabbit igg ba 1000, by Vector Laboratories (2 mentions)
Dylight 488 conjugated goat anti mouse igg, by Jackson ImmunoResearch (2 mentions)
Rabbit anti phospho smad1 5 8, by Cell Signaling Technology (1 mentions)
Dylight 488 conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Eclipse 800, by Nikon (1 mentions)
Fitc conjugated rat anti mouse igg2b, by BD (1 mentions)
3 protocols using dylight 549 conjugated goat anti rabbit igg
1
Immunofluorescent Staining of ISH-Stained Tissues
2
Immunofluorescent Staining of ISH-Stained Tissues
3
Colon Mucus Visualization Protocol
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Colon segments with contents maintained were fixed in Methanol–Carnoy's fixative according to a published protocol54 (link) with a few modifications. After 4 h of fixation in Methanol–Carnoy's, samples were incubated for 2 × 30 min in methanol, 2 × 20 min in ethanol, 2 × 25 min in xylene and 2 × 30 min in liquid paraffin before paraffin embedding.
To visualize intestinal mucus, 6 μm colon sections were stained for 1 h with polyclonal rabbit anti-mouse Muc2 (1:50, sc-15334, Santa Cruz), and mouse serum (1:50) collected 21 days after systemic priming with E. coli. After three washes in histological buffer (0.9 M NaCl, 20 mM Tris-HCl, pH 7.2), sections were incubated with DyLight549-conjugated goat anti-rabbit IgG (1:100, 111-505-144, Jackson Immunoresearch), FITC-conjugated rat anti-mouse IgG2b (1:100, 553395, BD) and 0.5 μg per ml DAPI (4′,6′-diamidino-2-phenylindole) and imaged using Nikon Eclipse 800 fluorescence microscope.
To visualize intestinal mucus, 6 μm colon sections were stained for 1 h with polyclonal rabbit anti-mouse Muc2 (1:50, sc-15334, Santa Cruz), and mouse serum (1:50) collected 21 days after systemic priming with E. coli. After three washes in histological buffer (0.9 M NaCl, 20 mM Tris-HCl, pH 7.2), sections were incubated with DyLight549-conjugated goat anti-rabbit IgG (1:100, 111-505-144, Jackson Immunoresearch), FITC-conjugated rat anti-mouse IgG2b (1:100, 553395, BD) and 0.5 μg per ml DAPI (4′,6′-diamidino-2-phenylindole) and imaged using Nikon Eclipse 800 fluorescence microscope.
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