Journal: Oncology Reports

Article Title: Use of CA15-3 for screening breast cancer: An antibody-lectin sandwich assay for detecting glycosylation of CA15-3 in sera

doi: 10.3892/or.2018.6433

Figure Lengend Snippet: Comparison of the levels of CA15-3 between groups in the sandwich ELISA. Mouse anti-CA15-3 monoclonal antibody (50 ng/well) and rabbit anti-CA15-3 polyclonal antibody (1:3,000 dilution ratio) were used as capture and detection antibodies, respectively. (A) The linear response of the ELISA as a function of serum dilution. The serum used for panel A was a mixture of stage III breast cancers (n=24). Serum (X) and Coating Ab (X) represent the results obtained when the serum and coating antibody reaction, respectively, were omitted. (B) The values obtained in the sandwich ELISA when a 1:100 serum dilution was used. Data are expressed as the means ± SEMs.

Article Snippet: Then the plates were reacted with serum samples (1:100 dilution ratio) prepared in 0.5% skim milk in PBS-T at 37°C for 2 h. The captured CA15-3 was detected using rabbit anti-MUC1 polyclonal antibody (1:3,000 dilution ratio; reaction: 37°C for 1 h; Sino Biological, Inc., Beijing, China; cat. no. 12123-T24), together with HRP-conjugated anti-rabbit IgG polyclonal antibody (1:5,000 dilution ratio; reaction: 37°C for 40 min, Bethyl Laboratories, Inc., Montgomery, TX, USA; cat. no. A120-100P).

Techniques: Sandwich ELISA, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Guishaozichuan granules can attenuate asthma in rats via the MUC5AC/EGFR signaling pathway

doi: 10.3389/fphar.2022.1011751

Figure Lengend Snippet: Effects of GSZC granules on lung MUC5AC (A) , EGFR (B) mRNA expression in rats suffering from asthma. Values are the mean ± SD, n = 6 for each group. ▲▲ p < .01 versus the normal group, ** p < .01 versus the model group.

Article Snippet: MUC5AC antibody was purchased from Boster Group (20180824; Wuhan, China).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Guishaozichuan granules can attenuate asthma in rats via the MUC5AC/EGFR signaling pathway

doi: 10.3389/fphar.2022.1011751

Figure Lengend Snippet: Effects of GSZC granules on lung MUC5AC (A) , EGFR (B) protein expression in rats suffering from asthma. Values are the mean ± SD, n = 6 for each group. ▲▲ p < .01 versus the normal group, * p < .05, ** p < .01 versus the model group.

Article Snippet: MUC5AC antibody was purchased from Boster Group (20180824; Wuhan, China).

Techniques: Expressing

Journal: Immunity, Inflammation and Disease

Article Title: Polydatin has anti‐inflammatory and antioxidant effects in LPS‐induced macrophages and improves DSS‐induced mice colitis

doi: 10.1002/iid3.455

Figure Lengend Snippet: PD maintained proper intestinal epithelial barrier in DSS‐induced mice. (A–C) Immunohistochemistry analysis the expression of Claudin‐1, Occludin and ZO‐1 of colon tissue from DSS and PD treatment mice, black arrow mark the protein ( n ≥ 4). (D, E) Immunohistochemistry analysis the expression of MUC2 and MUC3A in colon tissue from DSS and PD treatment mice, black arrow mark the protein ( n ≥ 4). (F) Mucicarmine staining analysis the mucus in DSS and PD treatment mice colon tissue, magnification is ×20, black arrow mark the mucus ( n ≥ 4). DSS, dextran sodium sulfate; PD, polydatin

Article Snippet: Primary antibodies zona occludens 1 (ZO‐1) (1:500; Proteintech), occludin (1:500; Abclonal), claudin 1 (1:500; Abclonal), mucin 2 (MUC2) (1:500; Proteintech), and mucin 3A (MUC3A) (1:500; Boster) were used to incubate the slides at 4°C overnight.

Techniques: Immunohistochemistry, Expressing, Staining

Journal: Scientific Reports

Article Title: Dietary polyamines promote intestinal adaptation in an experimental model of short bowel syndrome

doi: 10.1038/s41598-024-55258-4

Figure Lengend Snippet: Polyamines enhance mucosal defense factors in rats with massive intestinal resection. (A-B) Fecal and serum secretory IgA was measured using ELISA. n = 5–7/group. (C) IgA in ileum tissue was assessed using western blotting. n = 4–6/group. (D) Fecal mucin was measured using a fluorometric assay. n = 4–6/group. (E) Representative images of ileal villus showing immunostaining by Muc2 (original magnification, ×400; scale bars = 200 μm). Left graphs show the number of goblet cells per unit villous area and the size of goblet cell secretion granule. (F) The expression of Claudin-3 in the ileum tissue was measured by western blot analysis. Representative images of ileal villus showing immunostaining by Claudin-3 (original magnification, ×200; scale bars = 100 μm). (G) Serum DAO was measured by ELISA. n = 5–7/group. (H) Serum GLP-2 was measured by ELISA. n = 5–7/group. (I) Fecal short-chain fatty acid (SCFA) content was measured using high-performance liquid chromatography. n = 3–6/group. Data are presented as mean ± SD. Results of one-way ANOVA are represented as follows: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Nonspecific antibody binding was blocked using 15% goat serum for 30 min. For staining of Ki-67, Muc2, and Claudin-3, the slides were then incubated with anti-Ki67 antibody (ab16667; Abcam, Cambridge, UK), anti-Muc2 rabbit monoclonal antibody (M01212-1; Boster Biological Technology, Pleasanton, CA, USA), and Claudin-3 polyclonal antibody (#PA5-32353; Invitrogen, Waltham, MA, USA) at 4°C overnight, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Immunostaining, Expressing, High Performance Liquid Chromatography

Journal: Journal of Infection and Public Health

Article Title: A three-dimensional A549 cell culture model to study respiratory syncytial virus infections

doi: 10.1016/j.jiph.2020.03.011

Figure Lengend Snippet: RSVA infection and mucin production in A549 spheroids. A549 Spheroids were inoculated with 1 MOI of RSVA 2001/2-20 and incubated for 3 (3A) and 7 (3B ) days. At the assigned time-points, spheroids were immunostained with anti-RSV F antibody (green), anti-MUC1 antibody (red), and counterstained with the nuclear stain DAPI (blue) and visualized by confocal microscopy at 10× magnification (n = 3). Syncytial formation in RSV infected spheroids at day 3 post-infection was also shown in insets at 40× magnification. (3C) Mucin accumulation and (3D) RSV-F dissemination were measured by the Zeiss LSM710 laser scanning confocal microscope. Mucin intensity fold change was evaluated between infected spheroids and their time matching controls (n = 3). RSV-F intensity fold change was evaluated between day 3 and day 7 infected spheroids (n = 3). Error bars represent ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: After 3 and 7 days post-inoculation, spheroids were fixed with 4% paraformaldehyde (416785000 ACROS) for 15 min and permeabilized with 0.2% Triton X-100 (T9284-100 ml Sigma) in PBS for 20 min. Spheroids were blocked with blocking solution [1% BSA (A3912-100 g Sigma) and 0.1% Triton X-100 in PBS] for 1 h followed by overnight incubation with a mixture of mouse anti-RSV fusion protein antibody (MAB8599-Merck Millipore) at 1:200 and rabbit anti-human MUC1 antibody (12123-T24 Sino Biological) at 1:2500 in blocking buffer.

Techniques: Infection, Incubation, Staining, Confocal Microscopy, Microscopy