For quantification of metastatic lesions, mouse lungs were isolated, washed briefly in PBS, fixed in Bouin’s fixative (Sigma) overnight at 4 °C, and stored in 70% ethanol before counting lung nodules. For histological analysis, lungs were fixed in 10% formalin overnight at 4 °C, dehydrated, and embedded in paraffin (Tissue Tek Embedding Station). 3μm sections were cut on a Leica RM2255 rotary microtome. For antigen retrieval, deparaffinized slides were immersed in R-buffer A (Electron Microscopy Services) and boiled in a microwave for 20 min. Samples were blocked and permeabilized for 30 min in blocking buffer (5% normal goat serum, 0.3% TritonX-100 in PBS), followed by primary antibody incubation overnight at 4 °C using anti-CDH1 (1:100, BD Biosciences, 610181). Secondary antibody (1:500, BD Biosciences) was incubated for 1 h at RT. All sections were analyzed using a Nikon A1 confocal microscope and Zeiss fluorescence microscope.
Anti cdh1
Manufactured by BD
Sourced in United States
Anti-CDH1 is a laboratory reagent used for the detection and analysis of the CDH1 protein, which is involved in cell-cell adhesion. It is a tool used in research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Abcam (3 mentions)
Anti iκb, by Cell Signaling Technology (2 mentions)
Nitrocellulose membrane, by Merck Group (2 mentions)
Irdye 700 or 800 conjugated affinity purified secondary antibodies, by Rockland Immunochemicals (2 mentions)
Odyssey infrared imaging system, by LI COR (2 mentions)
P ikk, by Cell Signaling Technology (2 mentions)
P ikk, by Vector Laboratories (2 mentions)
Dylight conjugated secondary antibody, by Jackson ImmunoResearch (2 mentions)
Anti esr1, by Santa Cruz Biotechnology (2 mentions)
Rb 9017 p0, by Thermo Fisher Scientific (2 mentions)
Anti pgr, by Thermo Fisher Scientific (2 mentions)
P stat3, by Vector Laboratories (2 mentions)
P stat3, by Cell Signaling Technology (2 mentions)
Mmp 9, by Cell Signaling Technology (2 mentions)
P akt, by Cell Signaling Technology (2 mentions)
Vectastain elite abc kit, by Vector Laboratories (2 mentions)
Anti cd31, by Abcam (2 mentions)
Anti ki67, by BD (2 mentions)
Rm2255 rotary microtome, by Leica (1 mentions)
Bouin s fixative, by Merck Group (1 mentions)
7 protocols using anti cdh1
1
Quantification and Histological Analysis of Lung Metastases
2
Western Blot Analysis of Protein Targets
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Ten micrograms of total protein from the cell lysates were separated on SDS-PAGE gels and transferred to nitrocellulose membranes (Millipore Corp., Bedford, MA, USA). Membranes were blocked and incubated overnight with primary antibodies: anti-IκB (1:500 dilution, 9242, Cell Signaling Technology), anti-CDH1 (1:1000 dilution, 610181, BD Biosciences), and anti-β-actin (1:500 dilution, ab8229, Abcam). Immunoreactivity was visualized with IRDye 700 or 800 conjugated affinity-purified secondary antibodies (1:10000 dilution, Rockland Immunochemicals, Gilbertsville, PA, USA) using the Odyssey infrared imaging system (Li-COR, Lincoln, NE, USA).
3
Western Blot Analysis of Protein Targets
4
Comprehensive Protein Expression Analysis
5
Immunolocalization of Uterine Markers
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Immunolocalization of Ki67, CD31, ESR1, PGR, p-IKK, p-STAT3, CD68, CD163, COX2, iNOS, MMP9 and p-AKT was determined in cross-sections (5 μm) of paraffin-embedded uterine sections using specific primary antibodies and a Vectastain Elite ABC Kit (Vector laboratories, Burlingame, CA, USA) or DyLight-conjugated secondary antibody (Jackson ImmunoResearch Lab, West Grove, PA, USA). Antibodies used in these analyses were: anti-CDH1 (1:120 dilution, 610181, BD Biosciences, San Jose, CA, USA), anti-Ki67 (1:200 dilution, 550609, BD Biosciences), anti-CD31 (1:100 dilution, ab28364, Abcam, Cambridge, MA, USA), anti-ESR1 (1:100 dilution, sc-542, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-PGR (1:200 dilution, RB-9017-P0, Thermo Scientific, Rockford, IL, USA), p-IKK (1:150 dilution, 2697, Cell Signaling Technology, Danvers, MA, USA), p-STAT3 (1:50 dilution, 9145, Cell Signaling Technology), CD68 (1:300 dilution, ab955, Abcam), CD163 (1:300 dilution, ab126756, Abcam), COX2 (1:50 dilution, RM-9121, Thermo Scientific), iNOS (1:50 dilution, 610333, BD Biosciences), MMP9 (1:50 dilution, 3852, Cell Signaling Technology), and p-AKT (1:200 dilution, 3787, Cell Signaling Technology).
6
Western Blot Analysis of EMT Markers
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SDS lysis buffer (0.05 mM Tris-HCl, 50 mM BME, 2% SDS, 0.1% Bromophenol blue, 10% glycerol) was used to collect protein from cells. Samples were heat denatured. Protein was equally loaded, separated on a 10% SDS-page gel, transferred onto a pure nitrocellulose membrane (BioRad), and blocked with 5% milk. Primary antibodies for immunoblotting included anti-pSMAD2 (1:1000, Cell Signaling, 3108), anti-pSMAD3 (1:250, ThermoFisher, 44-246 G), anti-SMAD2/3 (1:1000, Cell Signaling, 3102), anti-CDH1 (1:5000, BD Biosciences, 610181), anti-CDH2 (1:2000, BD Biosciences, 610920), anti-ZEB1 (1:3000, Bethyl, A301-922A), anti-Vimentin (1:5000, BD Biosciences, 550513), and anti-β-Actin (1:5000, Abcam, ab8227) for loading control. Membranes were incubated with horseradish peroxidase (HRP)-conjugated anti-mouse secondary antibody (1:5000, GE Healthcare) or anti-rabbit secondary antibody (1:5000, GE Healthcare) for 1 h, after which chemiluminescent signals were detected using ECL substrate (GE Healthcare).
7
Immunolocalization of Uterine Markers
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