CORM-3 was purchased from Sigma Japan Inc. (Tokyo, Japan). CORM-3 solution was prepared by dissolving the compound in distilled water when it is in need. Meanwhile, iCORM-3 is prepared by dissolving CORM-3 in a phosphate buffer and standing to release CO at room temperature for 48 hours. Eliminate residual CO by bubbling the previous solution with N2 [20 (link)]. The following reagents were purchased from designated sources: anti-E-cadherin rabbit polyclonal antibody (cat no. 20874-1-AP), anti-N-cadherin rabbit polyclonal antibody (cat no. 22018-1-AP), anti-Keap-1 rabbit polyclonal antibody (cat no. 10503-2-AP), and anti-Nrf2 rabbit polyclonal antibody (cat no. 16396-1-AP) were purchased from Proteintech Group, Inc. (Chicago, IL, USA). Anti-HO-1 rabbit polyclonal antibody (cat no. ab13243) was the product of Abcam (Cambridge, MA, USA). Anti-GAPDH rabbit monoclonal antibody, anti-rabbit IgG secondary antibody, and anti-Ki67 rabbit monoclonal antibody were from Cell Signaling Technology (Beverly, USA).
Rabbit anti e cadherin polyclonal antibody
Manufactured by Proteintech
Sourced in United States
The Rabbit anti-E-cadherin polyclonal antibody is a laboratory reagent used to detect the expression of the E-cadherin protein in biological samples. E-cadherin is a cell adhesion molecule that plays a crucial role in maintaining cell-cell contacts and tissue integrity. This antibody can be used in various applications, such as Western blotting, immunohistochemistry, and immunofluorescence, to study the expression and localization of E-cadherin in different cell types and tissues.
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Lab products found in correlation
Fluo 4 am, by Thermo Fisher Scientific (2 mentions)
Anti rabbit igg secondary antibody, by Cell Signaling Technology (1 mentions)
Corm 3, by Merck Group (1 mentions)
Rabbit monoclonal anti gapdh antibody, by Cell Signaling Technology (1 mentions)
Anti ki67 rabbit monoclonal antibody, by Cell Signaling Technology (1 mentions)
Rabbit anti opn polyclonal antibody, by Proteintech (1 mentions)
Rabbit anti zeb1, by Proteintech (1 mentions)
Rabbit anti vimentin, by Cell Signaling Technology (1 mentions)
Anti n cadherin, by Cell Signaling Technology (1 mentions)
Giemsa staining, by Merck Group (1 mentions)
Fitc labeled goat anti rabbit igg, by Merck Group (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Proteintech (1 mentions)
Rabbit anti zo 1 polyclonal antibody, by Proteintech (1 mentions)
Rabbit anti gapdh polyclonal antibody, by Proteintech (1 mentions)
5 protocols using rabbit anti e cadherin polyclonal antibody
1
CO-Releasing Molecules Biological Characterization
2
Immunofluorescent Staining of E-cadherin
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Coverslips were placed horizontally on the bottom of a 6-well plate upon cleaning, disinfection and 24-h ultraviolet irradiation. Then, the cells were seeded on the coverslips at a density of 1x10 6 cells per well and cultured in an incubator. The coverslips were rinsed with PBS for 5 min 3 times and fixed with 4% paraformaldehyde for 15 min, followed by permeabilization of the cells with 0.3% Triton X-100 for additional 10 min. Next, the coverslips were rinsed with PBS again for 5 min 3 times and blocked by incubating the cells in 5% BSA for 60 min. Then, the cells were stained with anti-E-cadherin rabbit polyclonal antibody (dilution 1:100; ProteinTech Group, Inc.), followed by a 12-h incubation period at 4˚C. After washing the unbound antibody with PBS for 5 min 3 times, the cells were incubated with an Alexa 488-conjugated goat anti-rabbit immunoglobulin G secondary antibody (1:1,000; Cell Signaling Technology, Inc.) for 1 h at 4˚C in the darkness. Finally, DAPI was used as a counterstain to label the nuclei. The stained cells were then acquired and photographed with a fluorescence microscope.
3
Cancer Cell Response to CAF Secretome
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A total of 1.5 × 105 cells (MCF-7) or 1.7 × 105 cells (MDA-MB-231) were seeded on 6-well plates. After 24 h, the medium was replaced with CAFs CM for 72 h of incubation. Next, whole-cell extracts were prepared using NP-40 lysis buffer following the previously described protocol. Western blot experiments were performed on 20 μg protein samples using the protocol described above. The following primary antibodies (with respective dilutions) were used: rabbit anti-E-cadherin polyclonal antibody (1:5000), rabbit anti-OPN polyclonal antibody (1:1000), and rabbit anti-ZEB1 (zinc finger E-box binding homeobox 1) polyclonal antibody (1:1000), all from Proteintech, Rosemont, IL, USA. Densitometry analysis was carried out in ImageJ software with the tested protein levels normalized to β-actin and the respective untreated control (cancer cells untreated with CM).
4
Optimizing Cell Signaling Assays
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Recombinant human FST288 was purchased from R&D systems (Minneapolis, MN, USA). Cell Counting Kit-8 (CCK-8) was bought from GlpBio Biotechnology Co. (Shanghai, China). Rabbit anti-GAPDH polyclonal antibody and JNK inhibitor AS601245 were provided by Absin (Shanghai, China). Rabbit anti-vimentin, anti-N-cadherin, and anti-ezrin polyclonal antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Rabbit anti-E-cadherin polyclonal antibody was purchased from Proteintech (Chicago, IL, USA). Rabbit anti-JNK and anti-p-JNK antibodies were provided by Wanlei (Shenyang, China). Rabbit anti-MMP2, anti-p-Smad3 and anti-Smad3 polyclonal antibodies were provided by Abclonal (Woburn, MA, USA). Fluo-4 AM was purchased from Thermo Fisher Scientific (Ottawa, ON, Canada). Giemsa staining and FITC-labeled goat anti-rabbit IgG were provided by Sigma-Aldrich (Oakville, ON, Canada).
5
Western Blot Analysis of Rat Bladder
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For Western blot analysis, the rat bladder samples were processed according to the standard protocol (Beyotime, Shanghai, China). Total protein was extracted and quantified using the bicinchoninic acid (BCA) protein assay method. Approximately 40 μg of protein from each sample was separated on 8%, 10%, or 15% SDS-PAGE gels and transferred to polyvinylidene difluoride (PVDF) membranes. The membranes were then blocked with defatted milk and incubated overnight at +4°C with specific primary antibodies, including rabbit anti-E-Cadherin polyclonal antibody (1:5,000, Proteintech, United States), rabbit anti-ZO-1 polyclonal antibody (1:5,000, Proteintech, United States), rabbit anti-SNAP-23 polyclonal antibody (1:2000, Abcam, United States), mouse anti-cleaved and uncleaved SNAP-25 monoclonal antibody (1:100, GeneTex, United States), and rabbit anti-GAPDH polyclonal antibody (1:5,000, Proteintech, United States). Subsequently, the membranes were incubated with horseradish peroxidase-conjugated secondary antibody (1:3,000, Proteintech, United States) at room temperature for 1 h. Protein bands were visualized and analyzed using the Bio-Rad ChemiDoc Imagers System.
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