Rabbit anti muc2

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Rabbit anti-MUC2 is a primary antibody that specifically recognizes the MUC2 protein. MUC2 is a secreted mucin glycoprotein that plays a role in the protection and lubrication of mucosal surfaces. The Rabbit anti-MUC2 antibody can be used for the detection and study of MUC2 in various applications.

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Lab products found in correlation

Rabbit anti lysozyme, by Agilent Technologies (3 mentions) Rabbit anti ki67, by Thermo Fisher Scientific (1 mentions) Mouse anti brdu, by Thermo Fisher Scientific (1 mentions) Biotinylated donkey anti rabbit igg, by Vector Laboratories (1 mentions) Donkey anti mouse igg, by Vector Laboratories (1 mentions) Avidin horseradish peroxidase conjugates, by Vector Laboratories (1 mentions) 3 3 diaminobenzidine (dab), by Merck Group (1 mentions) Goat anti chromogranin a, by Santa Cruz Biotechnology (1 mentions) Fv1200 laser scanning microscope, by Olympus (1 mentions) Fv1200 confocal microscope, by Olympus (1 mentions) Click it edu kit, by Thermo Fisher Scientific (1 mentions) Rabbit anti chromogranin a, by Abcam (1 mentions) Tsa superboost kit, by Thermo Fisher Scientific (1 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (1 mentions) Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Biotinylated anti rabbit secondary antibody, by Santa Cruz Biotechnology (1 mentions) Szx16, by Olympus (1 mentions) Mouse anti e cadherin, by BD (1 mentions) Alexa fluor 488 and 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Mouse anti-Muc2 and rabbit anti-chromogranin A Anti-Muc2 Rabbit anti-TM

7 protocols using rabbit anti muc2

Immunohistochemical Analysis of Intestinal Markers

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Tissue sections were immunostained as previously described [28 (link)]. Primary antibodies included rabbit anti-Ki67 (Thermo Fisher Scientific, Inc., Fremont, CA; Cat. No. RM-9106-S1) (1:200), mouse anti-BrdU (Thermo; Cat. No. MS-1058-PO) (1:250), goat anti-cryptdin related sequence 4C (CRS4C) (gift from Dr. A. J. Ouellette, University of Southern California, Los Angeles, CA) [32 (link)] (1:2000), and rabbit anti-MUC2 (Santa Cruz Biotechnology, Inc, Santa Cruz, CA; Cat. No. sc15334) (1:100). Secondary antibodies included biotinylated donkey anti-rabbit IgG, donkey anti-goat IgG, and donkey anti-mouse IgG (all from Vector Labs, Burlingame, CA). Biotinylated antibodies were linked to avidin-horseradish peroxidase conjugates (Vector Labs), visualized using 3,3′-diamino benzidine (Sigma) for 2 to 5 min, and lightly counterstained with hematoxylin.

Aronson B.E., Stapleton K.A., Vissers L.A., Stokhuijzen E., Bruijnzeel H, & Krasinski S.D. (2014). Spdef deletion rescues the crypt cell proliferation defect in conditional Gata6 null mouse small intestine. BMC Molecular Biology, 15, 3.

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Immunofluorescence Analysis of Lgr5-GFP Organoids

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To monitor GFP expression, the Lgr5-GFP organoids were observed with Olympus FV1200 confocal microscope. For immunofluorescence, cultured organoids were first fixed for 1 h with 4% paraformaldehyde at room temperature. Organoids were washed with PBS in the eppendorf tubes for 3 times. Samples were permeabilized with 2% Triton X-100 for 2 h in the 4 °C and blocked with PBT solution (3%BSA and 1% Triton X-100 in PBS) for 2 h at room temperature. The organoids were then incubated overnight with the primary antibody at 4 °C. The following primary antibodies were used: rabbit anti-lysozyme (Dako, F0372, 1:200), rabbit anti-Muc2 (Santa Cruz, sc-15334, 1:300), goat anti-Chromogranin A (Santa Cruz, sc-1488, 1:300). The fluorescein-labeled secondary antibodies (Life Technologies, 1:300) and 4′, 6-diamidino-2-phenylindole (DAPI) were applied for 1 h at room temperature. 5-Ethynyl-2-deoxyuridine (EdU) staining was performed by following the manufacturer’s instruction (Click-IT; Invitrogen). Images were obtained with an Olympus FV1200 Laser Scanning Microscope.

Li Y., Liu Y., Liu B., Wang J., Wei S., Qi Z., Wang S., Fu W, & Chen Y.G. (2018). A growth factor-free culture system underscores the coordination between Wnt and BMP signaling in Lgr5+ intestinal stem cell maintenance. Cell Discovery, 4, 49.

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Comprehensive Intestinal Tissue Analysis

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Paraffin sections (4–5 μm) were stained with periodic acid-Schiff/Alcian blue (Newcomer Supply, cat. no. 9162B, 1003A) to visualize mucin-containing goblet cells. Immunostaining with rabbit anti-lysozyme (1:200, DAKO, cat. no. A0099), rabbit anti-GFP (1:200, Invitrogen, cat. no. A21311), rabbit anti-cleaved caspase-3 (1:50, Cell Signal, cat. no. 9664S), rabbit anti-Muc2 (1:200, Santa Cruz, cat. no. sc-15334), rabbit anti-chromogranin A (1:200, Abcam, cat. no. ab15160), and goat anti-DPP4 (1:200, Millipore, cat. no. SAB2500328) was performed as described (Lopez-Diaz et al., 2006 (link)). Co-staining for cleaved caspase-3 and MMP7 was performed by co-incubating rabbit anti-cleaved caspase-3 with rat anti-MMP7 (1:400, Vanderbilt, cat. no. 4,334). Rabbit anti-cleaved NOTCH1 (NICD; 1:50, Cell Signal, cat. no. 4147S) staining used a TSA SuperBoost kit (Thermo, no. B40943). EdU-Click-iT kit (Life Technologies, cat. no. C10337) was used to identify proliferating cells. Images were captured on a Nikon E800 microscope with Olympus DP controller software.

Bohin N., Keeley T.M., Carulli A.J., Walker E.M., Carlson E.A., Gao J., Aifantis I., Siebel C.W., Rajala M.W., Myers MG J.r., Jones J.C., Brindley C.D., Dempsey P.J, & Samuelson L.C. (2020). Rapid Crypt Cell Remodeling Regenerates the Intestinal Stem Cell Niche after Notch Inhibition. Stem Cell Reports, 15(1), 156-170.

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Immunohistochemistry of Cell Markers

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Immunohistochemistry was performed using standard protocols. Primary antibodies were incubated at 4°C overnight. Primary antibodies used were rabbit anti-pHH3 (Ser10) (Cell Signaling Technology #9701S, 1:500 dilution), rabbit anti-MUC2 (Santa Cruz Biotechnology #sc-15334, 1:500 dilution), rabbit anti-REG4 (abcam #ab255820, 1:500 dilution). Secondary antibodies (1:200 dilution) included a goat anti-rabbit Alexa Fluor 555 (ThermoFisher Scientific, #A27039).

Sharifkhodaei Z., Liu C.Y., Girish N., Huang Y., Punit S., Washington M.K, & Polk D.B. (2023). Colitis-induced upregulation of tumor necrosis factor receptor-2 (TNFR2) terminates epithelial regenerative signaling to restore homeostasis. iScience, 26(10), 107829.

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Histological and Immunohistochemical Analysis of Intestinal Development

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The intestinal tissues were fixed in 4% formalin solution, embedded in paraffin, and cut into 4-μm slices. After deparaffinization and hydration, H&E staining was performed for evaluation of intestinal development and inflammation. The intestinal development was assessed by measurement of villus/crypt length. Three hundred well-orientated villi/crypts randomly chosen from × 200 images was observed and measured under light microscope SZX16 (Olympus, Japan) for each mouse.
For immunohistochemistry, deparaffinized sections were incubated with primary antibodies rabbit monoclonal anti-Ki67 (ab16667, Abcam, Cambridge, MA, USA) and rabbit anti-MUC2 (Santa Cruz Biotechnology, Inc) overnight at 4°C. After washing in PBS, sections were further incubated with the biotinylated anti-rabbit secondary antibody (Santa Cruz Biotechnology, Inc) for 30 min followed by counterstaining with 3, 3′-diaminobenzidine. The expression of Ki67 and MUC2 was detected by counting the absolute number of positively stained cells. Five random areas from × 400 images were performed under light microscope DM5000 B (Leika, Germany) in at least 100 villi or crypts for each mouse.

Xie R., Sun Y., Wu J., Huang S., Jin G., Guo Z., Zhang Y., Liu T., Liu X., Cao X., Wang B, & Cao H. (2018). Maternal High Fat Diet Alters Gut Microbiota of Offspring and Exacerbates DSS-Induced Colitis in Adulthood. Frontiers in Immunology, 9, 2608.

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Immunostaining of Intestinal Enteroids

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On day eight post-crypt isolation, enteroids were fixed with 4% paraformaldehyde (PFA; Sigma-Aldrich) for 1 hour at 4 1C prior to permeabilization with 0.1% Triton X-100 (Sigma-Aldrich) and incubated in blocking buffer containing 10% goat serum (Sigma-Aldrich). Immunostaining was performed overnight at 4 1C using primary antibodies: mouse anti-E-cadherin (BD Transduction Laboratories), rabbit anti-muc2 (Santa Cruz) and rabbit anti-lysozyme (Dako), followed by Alexa Fluor-488 and -594 conjugated secondary antibodies (ThermoFisher Scientific). DNA was stained with DAPI (Molecular Probes). Images were acquired using a fluorescence microscope (Axioimager.M2, equipped with a Plan-Apochromat 63Â/1.4 oil immersion objective) and analysed using ImageJ/FIJI V1.51.

Treveil A., Sudhakar P., Matthews Z.J., Wrzesiński T., Jones E.J., Brooks J., Ölbei M., Hautefort I., Hall L.J., Carding S.R., Mayer U., Powell P.P., Wileman T., Di Palma F., Haerty W, & Korcsmáros T. (2020). Regulatory network analysis of Paneth cell and goblet cell enriched gut organoids using transcriptomics approaches. Molecular omics, 16(1).

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Immunofluorescence Protocol for Stomach Cells

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Human stomach sections were deparaffinized and submitted to antigen retrieval by hot steaming in the Target Retrieval solution (Dako North America, Inc., Carpinteria, CA). Cells grown on coverslips were fixed in 4% paraformaldehyde and then permeabilized by incubation in 0.3% Triton X-100 for 15 minutes at room temperature. Blocking for tissue section was performed during 1 h at RT in the serum-free protein blocking solution (Dako North America, Inc., Carpinteria, CA). Blocking for cells was performed during 1 h at room temperature in 10% donkey normal serum. The primary antibody incubation was performed in a humid chamber overnight at 4°C. Primary antibodies used were: 1) Mouse anti-HNF4γ (R&D Systems), at the concentration 1:1000 for tissue sections and 1:500 for cells; 2) Mouse anti-NR2F2 (Abcam), at the concentration of 1:500 and 3) Rabbit anti-MUC2 (Santa Cruz), at the concentration of 1:200. Appropriate secondary antibodies conjugated with Alexa 488, Cy3, or Cy5 were used (1h incubation at room temperature).

Sousa J.F., Nam K.T., Petersen C.P., Lee H.J., Yang H.K., Kim W.H, & Goldenring J.R. (2015). miR-30-HNF4γ and miR-194-NR2F2 regulatory networks contribute to the up-regulation of metaplasia markers in the stomach. Gut, 65(6), 914-924.

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