Anti aqp5

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-AQP5 is a laboratory reagent used in research applications. It is an antibody that specifically binds to the AQP5 protein, which is a water channel expressed in various tissues. The primary function of Anti-AQP5 is to facilitate the detection and study of AQP5 in biological samples.

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5 protocols using anti aqp5

Immunostaining of AQP5 in Embryos

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The in vivo and in vitro cultured embryos were fixed in 4% paraformaldehyde in PBS containing 3.7% picric acid in phosphate-buffered saline (PBS) for 30 min and then, washed PBS. Permeablization was conducted by 0.2% PBST (PBS containing 0.2% Triton X-100) for 3 hr. The embryos were blocked in 0.1% PBST containing 10% normal serum for 1hr at RT. After then, the embryos were incubated with rabbit polyclonal anti-AQP5 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a 1:200 dilution at 4°C overnight. Washed embryos were incubated with fluorescence 2nd antibody Cy^™3 conjugated AffiniPure (Jackson ImmunoResearch, USA) diluted 1:200 in PBS containing 2% BSA for 2 hr at RT. The nuclear of embryos was staining with Hochest33238 10 min. Dot slide were used for mounting. For negative controls, we deleted primary antibodies. Finally, fluorescent images were analyzed by Zeiss LSM 700 laser scanning microscopes with ZEN software.

Park J.W., Shin Y.K, & Choen Y.P. (2014). Adaptive Transition of Aquaporin 5 Expression and Localization during Preimplantation Embryo Development by In Vitro Culture. Development & Reproduction, 18(3), 153-160.

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Whole Mount Immunofluorescence of Embryos

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Whole mount immunofluorochemistry was followed previous report (Park et al., 2014 ). The in vivo collected and in vitro cultured embryos were fixed in 4% paraformal-dehyde in PBS containing 3.7% picric acid in phosphate-buffered saline (PBS) for 30 min. Permeabilzation was conducted by 0.2% PBST (PBS containing 0.2% Triton X-100) for 3 hr. The embryos were blocked in 0.1% PBST containing 10% normal serum for 1hr at RT. After then, the embryos were incubated with rabbit polyclonal anti-AQP5 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a 1:200 dilution at 4°C overnight. Embryos were incubated with fluorescence 2nd antibody Cy™3 conjugated AffiniPure (Jackson ImmunoResearch, USA) diluted 1:200 in PBS containing 2% BSA for 2 hr at RT. To stain the nuclear of embryos, Hochest33238 was used. Using dot slide, embryos were prepared for confocal imaging analysis. For negative controls, we deleted primary antibodies. Finally, fluorescent images were analyzed by Zeiss LSM 700 laser scanning microscopes with ZEN software

Park J.W, & Cheon Y.P. (2015). Temporal Aquaporin 11 Expression and Localization during Preimplantation Embryo Development. Development & Reproduction, 19(1), 53-60.

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Immunohistochemical Analysis of Lung Tissue

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All samples were cultured for 2 weeks, then fixed with 4% PFA for 2 days at 4 °C, followed by 20% sucrose in PBS for 12 h. Next, the samples were embedded in OTC compound, and sections (6 µm thick) were prepared using a cryostat (Thermo) and stained with hematoxylin-eosin (H&E) with a standard protocol. For immunohistochemistry, the sections were permeabilized with 0.1% Triton X-100 in PBS (TPBS) containing 1% BSA. The primary antibodies and dilutions in TPBS were as follows: anti-αtubulin (1:100, Santa Cruz), anti-TTF-1 (1:100, Santa Cruz), anti-SP-C (1:100, Santa Cruz), anti-AQP5 (1:100, Santa Cruz), and anti-CC10 (1:100, Santa Cruz). After incubation overnight at 4 °C and washing with TPBS 3 times, AlexaFluor 488 or 546 conjugated anti-goat, anti-rabbit, or anti-mouse secondary antibodies (Molecular Probes, Invitrogen) were used to detect primary antibodies. All nuclei were stained with DAPI (Dojin). Following 1 h of incubation at room temperature and washing with TPBS 3 times, fluorescence was examined using a fluorescence microscopy system (Zeiss Axiovert 200).

Kawai N., Ouji Y., Sakagami M., Tojo T., Sawabata N., Yoshikawa M, & Taniguchi S. (2018). Induction of lung-like cells from mouse embryonic stem cells by decellularized lung matrix. Biochemistry and Biophysics Reports, 15, 33-38.

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Immunostaining of Lung Cell Markers

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Cells were rinsed with DPBS and fixed with 4% paraformaldehyde (#P6148, Sigma-Aldrich) for 15 min. Samples were permeabilized with 0.5% Triton X-100 (#HFH10, Life Technologies) for 20 min and saturated in a blocking solution (3% bovine serum albumin (#A9056, Sigma-Aldrich), 0.1% Tween 20 (#P1379, Sigma-Aldrich), and 0.1% sodium azide (w/v) in DPBS) for 2 h at room temperature. The samples were further incubated with primary antibodies at 4 °C overnight, followed by the incubation with secondary antibodies in the blocking solution for 2 h and subsequent nuclei staining in PBS containing 1 μg/mL DAPI for 15 min at room temperature. The following primary antibodies were used for immunostaining: anti-NKX2.1 (1:100, #MA5-13961), anti-ZO-1 (1:100, #40-2200), anti-SFTPC (1:25, #PA5-71680), anti-ACE2 (1:1000, #MA5-31394) and anti-TMPRSS2 (1:50, #PA5-14264) from Invitrogen, anti-AQP5 (1:100, #SC-9890) from Santa-Cruz. All Alexa Fluor conjugated secondary antibodies are from Invitrogen, and the corresponding secondary antibodies were diluted in 1:250. All samples were observed with an LSM Zeiss 710 confocal microscope (Zeiss, France). Images were collected as TIF files and analysed with the software ImageJ.

Rofaani E., He Y., Peng J, & Chen Y. (2023). Epithelial folding of alveolar cells derived from human induced pluripotent stem cells on artificial basement membrane. Acta biomaterialia, 163.

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Molecular Mechanisms of Pilocarpine-Induced Pancreatic Injury

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Pilocarpine hydrochloride, streptozotocin (STZ), and citric acid were procured from Sigma Chemical Company (St. Louis, MO, USA). The following proteins were used in this study: antibodies against anti-amylase (#4017, Cell Signaling Technology, Danvers, MA, USA), anti-NHE-1 (sc-28758, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-AQP-5 (sc-514022, Santa Cruz Biotechnology, CA, USA), anti-GRP78 (sc-376768, Santa Cruz Biotechnology, CA, USA), anti-CHOP (#2895, Cell Signaling Technology, Danvers, MA, USA), p-IRE1α (ab48187, Abcam, Cambridge, MA, USA), IRE-1α (#3294, Cell Signaling Technology, Danvers, MA, USA), anti-p-eIF2α (#9721, cell signaling, Danvers, MA, USA), anti-eIF2α (sc-133132, Santa Cruz Biotechnology, CA, USA), and anti-β-actin (sc-130300, Santa Cruz Biotechnology, CA, USA). Horseradish peroxidase-conjugated secondary antibodies were obtained from Enzo Life Sciences, Inc. (Farmingdale, NY, USA).

Lee H.Y., Gu M., Cheng J., Suh J.W, & Chae H.J. (2020). Ixeris dentata and Lactobacillus gasseri Extracts Improve Salivary Secretion Capability in Diabetes-Associated Dry Mouth Rat Model. Nutrients, 12(5), 1331.

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