For mouse lung immunohistochemistry, the deparaffinized sections were stained with the following primary antibodies: rabbit anti-E-cadherin and rabbit anti- MMP7 (Cell Signaling Technology, Danvers, MA, USA), rabbit anti-SP-C (Hycult Biotech, Uden, Netherlands), and ACE-2 (R&D Systems, Minneapolis, MN, USA). For human lung immunohistochemistry, deparaffinized sections were stained with the following primary antibodies: rabbit anti-E-cadherin (Cell Signaling Technology), rabbit anti-proSP-C (Merck, Darmstadt, Germany), rabbit anti-ACE-2 (R&D Systems), and mouse anti-Laminin γ2 N-terminal fragment (γ2pf, Funakoshi, Tokyo, Japan). Histofine simple stain mouse MAX-PO secondary antibodies (Nichirei, Tokyo, Japan) and the Opal multiplex fluorescent immunohistochemistry system (Akoya Biosciences, Marlborough, MA, USA) were used according to the manufacturer’s protocol. All the images were captured using a fluorescence microscope (BZ-X710; Keyence, Osaka, Japan). To calculate the percentage of Ace2+ cells in E-cadherin+ bronchioles or invasive epithelial cells from the UIP lungs of four mice, five images (200× magnification) were captured and the percentage of Ace2 positive cells was calculated by ImageJ Fiji.
Rabbit anti e cadherin
Manufactured by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Germany
Rabbit anti-E-cadherin is a primary antibody that specifically recognizes the E-cadherin protein. E-cadherin is a calcium-dependent cell-cell adhesion glycoprotein that is important for the maintenance of epithelial cell layer integrity and polarity.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti vimentin, by Cell Signaling Technology (20 mentions)
Rabbit anti n cadherin, by Cell Signaling Technology (10 mentions)
Rabbit anti snail, by Cell Signaling Technology (8 mentions)
Mouse anti cd44, by Bio-Rad (5 mentions)
Triton x 100, by Merck Group (5 mentions)
Anti vimentin, by Cell Signaling Technology (4 mentions)
Rabbit anti gsk 3β, by Cell Signaling Technology (4 mentions)
Rabbit anti β1, by Abcepta (4 mentions)
Mouse anti fyn, by BioLegend (4 mentions)
Rabbit anti ki67, by Abcam (4 mentions)
Rabbit anti gapdh, by Cell Signaling Technology (4 mentions)
Mouse anti α tubulin, by Merck Group (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions)
Ripa buffer, by Thermo Fisher Scientific (4 mentions)
Rabbit anti β catenin, by Cell Signaling Technology (3 mentions)
Mouse anti albumin, by R&D Systems (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions)
Anti n cadherin, by Cell Signaling Technology (3 mentions)
Pvdf membrane, by Merck Group (3 mentions)
Rabbit anti mtor, by Cell Signaling Technology (3 mentions)
87 protocols using rabbit anti e cadherin
1
Mouse and Human Lung Immunohistochemistry
2
E-cadherin Immunoprecipitation Protocol
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Co-immunoprecipitation assay was performed as previously described (Sun et al., 2018 (link)). Cells were lysed in immunoprecipitation buffer supplemented with complete protease inhibitor. After lysing with ultrasound, lysates were centrifuged to remove insoluble components. Lysates were then incubated overnight with rabbit anti-E-cadherin (Cell Signaling Technology) or rabbit IgG primary antibodies at 4°C. Protein G beads (Beyotime) were then added to the lysates. The precipitated proteins were measured via western blot analysis as described above.
3
Immunofluorescence Staining of Cell Markers
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Cells were cultured on cover glasses in 24‐well cell culture plates, fixed with 4% PFA at room temperature for 10 min, and washed thrice with PBS. Rabbit anti‐E‐cadherin (Cell Signaling Technology, 3195, 1:200), mouse anti‐N‐cadherin (Invitrogen, 333900, 1:200), rabbit anti‐Nanog (Novus Biologicals, NB100‐58842, 1:300), and mouse anti‐Rex1 (made in‐house, 1:500) were added as the primary antibodies prepared with 0.3% Triton X‐100 and 10% goat serum in PBS at 4°C overnight. Then, cells were washed thrice with PBS. Goat anti‐rabbit Alexa 647 (Abcam, ab150079, 1:500) and goat anti‐mouse Alexa 568 (Invitrogen, A11004, 1:500) were added as the secondary antibodies prepared with PBS at room temperature for 1 h. Then, cells were washed thrice with PBS. Nuclei were stained with DAPI prepared in PBS at room temperature for 5 min. Image collecting was conducted with Zeiss LSM 800.
4
Western Blot Analysis of Epithelial-Mesenchymal Transition Markers
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The cultured cells were rinsed with cold PBS before treated with RIPA lysis buffer at 4 °C for 10 min. Then the mixture was heated at 100 °C for 10 min and centrifuged under 4 °C at 14000 g min−1 for 10 min. The supernatant was removed, and the protein concentration was measured with the BCA method. About 20 μg of protein was loaded in each lane, separated by 10% SDS–PAGE and transferred to the PVDF membrane. The membrane was blocked with 5% non-fat milk powder for 1 h at room temperature before overnight incubation with primary antibodies 4 °C, followed by the secondary antibody. The antibodies were mouse anti-β-tubulin (Cell Signaling, Danvers, MA, USA; CAT 6181), rabbit anti-MBD3 (Cell Signaling, CAT 3896), mouse anti-Flag (Sigma, San Francisco, CA, USA; CAT F1804), rabbit anti-MMP2 (ImmunoWay, Plano, TX, USA; CAT YT2798), rabbit anti-MMP9 (ImmunoWay CAT YT1892), rabbit anti-Vimentin (Cell Signaling, CAT 5741), rabbit anti-N-cadherin (Cell Signaling, CAT 13116), rabbit anti-E-cadherin (Cell Signaling, CAT 3195), rabbit anti-β-catenin (Cell Signaling, CAT 8480), rabbit anti-Snail (Cell Signaling, CAT 3879), rabbit anti-α-SMA (Cell Signaling, CAT 14968), rabbit anti-Smad2/3 (Cell Signaling, CAT 8685), rabbit anti-P-Smad2 (Cell Signaling, CAT 3108), rabbit anti-P-Smad3 (Cell Signaling, CAT 9520), rabbit anti-TGF-β (Cell Signaling, CAT 3709).
5
Analyzing Cell Signaling Pathways
6
Immunofluorescence Characterization of Stem Cells
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Immunofluorescence was performed as described.33 (link) The following primary antibodies and dilutions were used: rat anti-CD31 1 : 1000 (BD, Franklin Lakes, NJ, USA), mouse anti-Tuj-1 1 : 1000 (Covance, Princeton, NJ, USA), mouse anti-AFP 1 : 1000 (Inmunostep, Salamanca, Spain), rabbit anti-Nanog 1:1000 (Chemicon, Billerica, MA, USA), mouse anti-SSEA-1 (MC-480) 1 : 100 (Pierce, Waltham, MA, USA), rabbit anti-E-cadherin 1 : 60 (Cell Signaling, MA, USA), mouse anti-E-cadherin 1:200 (Cell Signaling, Danvers, MA, USA), rabbit anti-β-catenin 1 : 200 (BD). Secondary antibodies were: Alexa 647 goat anti-IgG rabbit (Molecular Probes, Eugene, OR, USA), Alexa 488 goat anti-IgG mouse (Molecular Probes), Alexa 568 donkey anti-IgG rat (Molecular Probes), Cy3 donkey anti-IgG rabbit (Jackson Immunoresearch, West Grove, PA, USA) and FITC donkey anti-IgG mouse (Jackson Immunoresearch). Images were obtained with NIKON EclipseTE2000 and ZEISS LSM 800 confocal microscope.
7
Immunofluorescence Protocol for Cellular Protein Analysis
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Cells were fixed with 4% paraformaldehyde for 15 min at room temperature, and then blocked with PBS containing 0.25% Triton X-100 (Sigma) and 3% BSA (Sigma) for 1 h at room temperature. The cells were then incubated with primary antibodies at 4 °C overnight, washed with PBS, and incubated with appropriate fluorescence-conjugated secondary antibody for 1 h at room temperature in the dark. Nuclei were stained with DAPI (Sigma). Primary and secondary antibodies were diluted in PBS containing 3% BSA. Images were captured at room temperature using an Olympus microscope (IX71) or a confocal microscope (TCS SP5; Leica) with a 63× oil objective. The p-ATM intensity was quantified by software LAS AF Lite (Leica). Antibodies used for immunofluorescence are as follows: rabbit anti-E-cadherin (Cell Signaling, 1:200), mouse anti-albumin (R&D, 1:200), mouse anti-p-ATM (Ser1981) (Thermo, 1:200), rabbit anti-HA-Tag (Cell Signaling, 1:200), rat anti-RPA32 (Cell Signaling, 1:200), rabbit anti-53BP1 (Novus, 1:200), goat anti-Nanog (R&D, 1:100), mouse anti-SSEA1 (R&D, 1:100), Cy3-conjugated donkey anti-mouse/rabbit/rat/goat IgG (Jackson Laboratories, 1:1 000), Cy5-conjugated goat anti-rabbit/mouse IgG (Jackson Laboratories, 1:1 000).
8
Immunohistochemical Analysis of Lung Development
9
Western Blot Protein Detection Protocol
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Cells were lysed with Whole Cell Lysis Assay kit (Keygen Biotech, China). Protein samples were electrophoresed in SDS-PAGE and then transferred to PVDF membranes (Millipore, USA). After blocked for 1 h with 5% skim milk at room temperature, the membranes were incubated with primary antibodies at 4℃ overnight. The membranes were washed thrice with TBST, and then incubated with secondary antibodies (Keygen Biotech, China). Signals were detected using an ECL system (Tanon, China) and quantified by ImageJ software. Mouse anti-NSD2 (Abcam, UK), rabbit anti-E-cadherin, anti-N-cadherin, anti-Vimentin (Cell Signaling Technology, USA) and rabbit anti-GAPDH (Keygen Biotech, China) were used in this study.
10
Immunohistochemical Analysis of Placental Tissues
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Placentas were fixed in 4% paraformaldehyde for 24–36 h following a standard protocol, then dehydrated and embedded in paraffin. Tissue sections (5-μm thickness) were mounted on glass slides (Thermo Scientific, Waltham, MA, USA). Rabbit anti-E-cadherin, anti-vimentin, and anti-β-cadherin polyclonal antibodies (1:100; Cell Signaling Technology) were used as primary antibodies, and anti-rabbit IgG (1:200; Sigma-Aldrich) used as a secondary antibody. Sections were mounted onto slides using Gel Mount Aqueous Mounting Medium (Sigma-Aldrich) and examined with an Olympus BX51 microscope.
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