alcian blue stain kit  (Vector Laboratories)

Journal: Gut Microbes

Article Title: Microbial metabolite Enterobactin impairs mitochondrial respiration and alleviates colitis

doi: 10.1080/19490976.2026.2662638

Figure Lengend Snippet: Monomeric Ent, 2, 3-DHBA, but not 2, 5-DHBA, restores mucosal integrity and mitochondrial homeostasis in DSS-induced colitis. Acute colitis was induced in mice as in Figure 7. After the cessation of DSS (recovery phase) mice were given either 2, 3-DHBA or 2, 5-DHBA (500 µg/mouse per day) for 7 d and euthanized. (A) Representative H&E-stained colon sections showing epithelial damage, crypt loss, and inflammatory cells ( n = 5/group). (B) Alcian blue staining for goblet cells in the colon (C) Immunoblot analysis of tight junction proteins (occludin and claudin-1) in colonic tissue. (D) qRT-PCR analysis of genes involved in mitochondrial biogenesis peroxisome proliferator activated receptor gamma coactivator 1-alpha ( PGC-1α ) and deiodinase type 2 ( Dio2) .(E) Colonic expression of antioxidant genes (Superoxide dismutase 2 (SOD2) , glutathione peroxidase (Gpx) , and nuclear factor erythroid 2-related factor 2 (Nrf2) (F) Immunoblot showing protein involved in mitochondrial dynamics uncoupling protein 1 (Ucp1), mitofusin 1 (Mfn1), and optic atrophy type 1 (Opa1). Data is presented as mean ± SEM ( n = 5). Statistical comparisons were performed using one-way ANOVA with Tukey’s post hoc test (* p < 0.05, ** p < 0.01, and *** p < 0.001).

Article Snippet: Alcian blue staining was performed to visualize acidic mucopolysaccharides and glycosaminoglycans in goblet cells of colon sections using the Alcian Blue Stain Kit (pH 2.5, Cat# H-3501, Vector Laboratories), following the manufacturer’s instructions.

Techniques: Staining, Western Blot, Quantitative RT-PCR, Expressing

Journal: STAR Protocols

Article Title: Protocol for developing a mucus-producing gut-on-a-chip model from Caco-2 and HT29-MTX-E12 cells

doi: 10.1016/j.xpro.2025.104321

Figure Lengend Snippet: Mucus production, barrier integrity and cell viability evaluation for GoC characterization (A) Mucus production in co-culture 9:1 and 7:3 with Alcian blue/Periodic Acid Schiff’s staining. (B) Barrier integrity with LY (20 μg/ml) for co-culture 9:1 and 7:3 day 2, 4, 6, 8; blank - empty chip (Data are represented as mean ± SD. Test: One-way ANOVA t test, p < 0.01 “∗∗”, p < 0.0001 “∗∗∗∗”, n = 4). (C) Barrier integrity with LY (20 μg/ml) for co-culture 9:1 and 7:3 day 2, 4, 6, 8 (Data are represented as mean ± SD. Test: Two-way ANOVA t test, p < 0.05 “∗”, non-significant “ns”, n = 4). (D) Cell viability with CCK-8 cell viability kit for co-culture 9:1, 7:3 and viability control for co-culture 7:3 (0.25% Triton X-100 added in CM in flow for 24 hours at 2 μL/min) (Data are represented as mean ± SD. Test: One-way ANOVA t test, non-significant “ns”, p < 0.0001 “∗∗∗∗”, n = 4). (E) Variance using barrier integrity test with LY (20 μg/ml) for co-culture 7:3 day 2, 4, 6, 8 (Data are represented as mean ± SD. Test: Two-way ANOVA t test, p < 0.05 “∗”, p < 0.0001 “∗∗∗∗”, non-significant “ns”, n = 12–3 independent chips). (F) Variance using cell viability test with CCK-8 cell viability kit for co-culture 7:3 on day 8 in three chips independently No1, No2, No3 (n = 4) and all of them combined No1 + No2 + No3 (n = 12) (Data are represented as mean ± SD. Test: One-way ANOVA t test, non-significant “ns”).

Article Snippet: EprediaTM Alcian Blue/PAS Special Stain Kit , Fisher Scientific , 23-900-664.

Techniques: Co-Culture Assay, Staining, CCK-8 Assay, Control

Journal: Nature Microbiology

Article Title: Weaning drives microbiome-mediated epigenetic regulation to shape immune memory in mice

doi: 10.1038/s41564-026-02295-6

Figure Lengend Snippet: ( a ) Representative images of IHC staining for Ki-67 and Alcian blue (AB) in colon tissues from mice under SPF (n = 4) or GF (n = 3) housing conditions. Scale bars, 50 μm. ( b ) Quantification of Ki-67 + cells and AB + goblet cells across intestinal segments (duodenum, jejunum, ileum, and colon) in SPF and GF mice. All data are presented as mean ± s.e.m. of at least 3 biological replicates. P values were calculated using unpaired (two-tailed) t -test.

Article Snippet: Alcian blue (AB) (pH 2.5) staining (Vector Labs, H-3501) was conducted following manufacturer instructions.

Techniques: Immunohistochemistry, Two Tailed Test