The proteins were extracted from the whole cell lysates using RIPA cell lysis buffer and the protein concentration was determined by the Bradford reagent. In total, 20 μg of the extracted total cellular protein from each sample were separated via SDS-PAGE, and transblotted onto EMD Millipore Immobilon™-P PVDF Transfer Membranes (EMD Millipore Cat No.: IPVH00010, USA). Western blot analyses were conducted with the following antibodies: mouse monoclonal anti-CD109 (Cat. No. 556039, BD Biosciences), anti-fibronectin (Cat. No. 610078, BD Biosciences), anti-Vimentin (D21H3) XP® Rabbit (Cat. No. 5741. Cell signaling); Anti-ZEB2 (Cat. No. sc-271984, Santa Cruz Biotechnology); rabbit polyclonal anti-Snail (Cat. No. ab5351, Abcam), anti-twist (Cat. No. ab505181; Abcam), anti-TGFBR1 (ALK5) (Cat. No. AHO1552, Invitrogen), anti-N Cadherin (Cat. No. ab18203, Abcam), anti-E-Cadherin (Cat. No. ab216783, Abcam), Anti-P-Smad2 (Cat. No. mAb #3108, cell signalling); Anti-P-Smad2/3 (Cat. No. MAB8935, R&D system, and anti- Slug (Cat. No. 9585S Cell signaling), anti-β-actin antibodies (Cat. No. sc-47778, Santa Cruz Biotechnology).
Anti n cadherin
Manufactured by Thermo Fisher Scientific
Sourced in United States
Anti-N-cadherin is a laboratory reagent used for the detection and quantification of N-cadherin protein in biological samples. N-cadherin is a cell adhesion molecule important for cell-cell interactions. Anti-N-cadherin can be used in various biochemical and immunological techniques to study N-cadherin expression and function.
Automatically generated - may contain errors
Lab products found in correlation
Anti e cadherin, by Thermo Fisher Scientific (5 mentions)
Anti snail, by Cell Signaling Technology (3 mentions)
Anti zo 1, by Thermo Fisher Scientific (3 mentions)
Anti myc, by Thermo Fisher Scientific (2 mentions)
Anti phospho smad2 3, by Cell Signaling Technology (2 mentions)
Anti smad2, by Cell Signaling Technology (2 mentions)
Anti cdk10, by Thermo Fisher Scientific (2 mentions)
Anti β catenin, by Fortis Life Sciences (2 mentions)
Lsm 700 confocal microscope, by Zeiss (2 mentions)
Anti e cadherin, by BD (2 mentions)
Anti e cadherin, by Abcam (2 mentions)
α tubulin, by Merck Group (2 mentions)
Anti β actin, by Thermo Fisher Scientific (2 mentions)
Anti cleaved caspase 3, by Thermo Fisher Scientific (2 mentions)
Anti cyclin d1, by Thermo Fisher Scientific (2 mentions)
Anti α tubulin, by Merck Group (2 mentions)
Anti α actinin, by Merck Group (2 mentions)
Anti β catenin, by BD (2 mentions)
Anti connexin 43, by Proteintech (2 mentions)
Anti desmoglein 2, by Abcam (2 mentions)
26 protocols using anti n cadherin
1
Comprehensive Protein Analysis in Cell Lysates
2
USP37 Regulation via Mutagenesis and Antibody Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human USP37 cDNA was subcloned into pcDNA3.1-Flag vector or PCDH vector and USP37-C350S was made using PCR-based site-directed mutagenesis method. All other constructs were generated using standard molecular cloning methods and were confirmed by DNA sequencing. Antibodies were commercially purchased: anti-USP37 (rabbit, 18465-1-AP, Proteintech), anti-Snail (rabbit, 3879, Cell Signaling), anti-Ubi (mouse, sc-8017, Santa Cruz), anti-actin (mouse, 60008-1-lg, Proteintech), anti-N-cadherin (mouse, 33-3900, Invitrogen), anti-HA (mouse, MMS-101P, Covance), anti-Flag (mouse, F3165, Sigma), anti-Flag (rabbit, F7425, Sigma), anti-Myc (mouse, 13-2500, Invitrogen), anti-GAPDH (mouse, 60004-1-Ig, Proteintech), normal IgG (rabbit, sc-2027, Santa Cruz), and GSH beads (GE). MG132 and cycloheximide (CHX) were obtained from Sigma.
3
Western Blot Analysis of EMT Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cell lysates and conditioned media were prepared, and protein concentrations were determined using a Bradford assay kit (Pierce Biotechnology, Rockford, IL). Equivalent amounts of proteins were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) on a 10% gel. Separated proteins were transferred to a nitrocellulose membrane (pH 7.9; Amersham Biosciences Inc., Piscataway, NJ), blocked with 5% non-fat powdered milk, and incubated with specific anti-LCN2 (R & D Systems; AF1757), anti-Twist (GeneTex; GTX127310), anti-Snail (Cell Signaling; #6615), anti-N-cadherin (Invitrogen; 33–3390) and anti-fibronectin (Santa Cruz Biotechnology; sc-9068) primary antibodies at 4°C overnight. After washing, membranes were incubated with HRP-conjugated anti-mouse, anti-rabbit or anti-goat IgG secondary antibody, as appropriate, for 1 h at room temperature. Immune complexes were visualized using an enhanced chemiluminescence (ECL) detection kit (Amersham) and Fuji X-ray film.
4
Western Blot Analysis of Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cultured cells were washed twice with ice-cold PBS. They were incubated with 2× SDS buffer (100 mM Tris-HCl [pH 6.8], 2% [w/v] SDS, 0.01% bromophenol blue, 20% glycerol, and 10% β-mercaptoethanol) for 5 min at 25 °C. The cells were then collected by scraping with cell scrapers and proteins were denatured at 95 °C for 5 min. Western blot analysis was performed using the following primary antibodies: anti‐phospho‐Smad 2/3 (1:1,000; Cell Signaling Technology, Danvers, MA, USA), anti‐Smad2 (1:1,000; Cell Signaling Technology), anti-N-cadherin (1:2,000, Invitrogen), anti‐phospho-ERKs (1:5,000; Cell Signaling Technology), anti‐ERKs (1:2,000; Cell Signaling Technology), anti‐phospho‐Akt (1:4,000; Cell Signaling Technology), anti‐Akt (1:1,000; Cell Signaling Technology), anti‐phospho‐p38 (1:1,000; Cell Signaling Technology), anti‐p38 (1:1,000; Cell Signaling Technology), anti-Smad4 (1:1,000; Cell Signaling Technology), anti-OB-cadherin (1:1,000; Cell Signaling Technology), and α‐tubulin (1:20,000; Sigma). Densitometry of the bands obtained was performed with the ImageJ software (NIH, Bethesda, MD, USA).
5
Immunohistochemical Staining of Tissue Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The IHC staining procedure was adopted. In a summarized protocol, paraffin-embedded tissue samples were sectioned into 4-μm slices. Following deparaffinization and rehydration, the sections underwent antigen retrieval using 1 mM EDTA (pH 8.0). To neutralize the tissue's inherent peroxidase activity, sections were treated with 0.3% hydrogen peroxide. Blocking was achieved using 5% goat serum. This was followed by an incubation phase with primary antibodies: anti-RNF115 (1:200), anti-CDK10 (1:200), anti-Ki67 (1:200), anti-E-cadherin (10 µg/mL), and anti-N-cadherin (1:100) sourced from Invitrogen, CA, USA. Subsequently, sections were incubated with biotinylated secondary antibodies and observed under a microscope (Olympus, Japan).
6
Western Blot Analysis of Protein Biomarkers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein lysates from tissues or cells were prepared in IP lysis buffer supplemented with protease inhibitor cocktail (Pierce, Thermo Fisher Scientific). Protein estimation was performed using the BCA Protein Assay Kit (Pierce). Equal amounts of protein lysates were resolved by SDS-PAGE and transferred onto polyvinylidene fluoride (PVDF) membrane for western blotting. Membranes were blocked with 5% non-fat milk, followed by incubation with primary antibody at 4°C overnight: anti-HOXA9 (ab140631, Abcam, Cambridge, UK), anti-β-catenin (71-2700, Invitrogen), anti-Twist (ab50581, Abcam), anti-E-cadherin (3195, Cell Signaling Technologies, Beverly, MA, USA), anti-N-cadherin (33-3900, Invitrogen), anti-Slug-1 (9585, Cell Signaling Technologies), anti-YAP1 (sc-15407, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-HIF-1α (sc-10790, Santa Cruz), anti-CTCF (ab70303, Abcam), and anti-GAPDH (sc-25778, Santa Cruz). Membranes were then incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody (1:5,000, Bioworld Technology, Louis Park, MN, USA) at room temperature for 1 h. Western blots were visualized using Immobilon Western Chemiluminescent HRP Substrate (Merck Millipore, Burlington, MA, USA) followed by film exposure. The relative expression of a target protein was normalized with internal control.
7
Protein Extraction and Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein extraction from tumor tissues and cells was performed using RIPA buffer, with the subsequent western blotting procedure carried out as detailed in earlier methods (Bacopulos et al. 2012 (link)). The primary antibodies used for western blotting were anti-RNF115 (0.4 µg/mL), anti-E-cadherin (1:10,000), anti-N-cadherin (1 µg/mL), anti-CDK10 (1:2,000), anti-SFN (2 µg/mL), anti-MYC (2 µg/mL), anti-p-Raf-1 (1:1,000), anti-Raf-1 (1:2,000), anti-p-MEK1/2 (1;1,000), anti-MEK1/2 (1:1,000), anti-p-ERK1/2 (1:1,000), anti-ERK1/2 (1 µg/mL), anti-CyclinD1 (1 µg/mL), anti-CDK4 (2 µg/mL), anti-Bax (1:100), anti-Cleaved caspase-3 (1:2,000), and anti-β-actin (1:5,000) (Invitrogen, CA, USA).
8
Western Blot Analysis of BM-MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BM-MSCs were washed twice with ice-cold PBS. They were then incubated with 2× SDS buffer (100 mM Tris-HCl (pH 6.8), 2% (w/v) SDS, 0.01% bromophenol blue, 20% glycerol, and 1.43 M β-mercaptoethanol) for 5 min at 25 °C. Then, the proteins were denatured at 95 °C for 5 min. Western blot analysis was performed with the following primary antibodies: Anti-phospho-Smad 2/3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), anti-Smad2 (1:1000; Cell Signaling Technology), anti-N-cadherin (1:2000; Invitrogen), and anti-α-tubulin (1:20,000; Sigma). Densitometry of the bands obtained was carried out with ImageJ software (NIH, Bethesda, MD, USA).
9
Immunostaining of Cell-Cell Junctions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary antibodies used for immunostaining were: anti-αE-catenin (1:100; Enzo Life Sciences ALX-804-101-C100), anti-β-catenin (1:250; BD Transduction Laboratories 610154), anti-plakoglobin (1:100; Cell Signaling 2309), anti-vinculin (1:800; Sigma Aldrich V9131), anti-N-cadherin (1:250; Invitrogen 99-3900), anti-l-afadin (1:500; Sigma Aldrich A0349), anti-connexin-43 (1:100; ProteinTech 15386-1-AP), anti-plakophilin 2 (1:10; Progen 651101), anti-desmoglein 2 (1:250; Abcam EPR6768), anti–α-actinin (1:250; Sigma A7811), anti-Mena (1:300, mouse monoclonal, a kind gift from Frank Gertler, Massachusetts Institute of Technology), and anti-Cre recombinase (1:300; Cell Signaling 12830). Secondary antibodies used were goat anti-mouse or anti-rabbit immunoglobulin G conjugated to Alexa Fluor-488, 568, or 647 (1:250; Invitrogen). F-actin was visualized using an Alexa Fluor dye conjugated to phalloidin (1:100; Thermo Fisher Scientific).
10
Immunodetection of Cell Adhesion Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rabbit polyclonal anti-TMEM43 (N-13) antibody (Santa Cruz, CA, USA) was used in immunofluorescence microscopy and immunoblot analysis to detect TMEM43. We used mouse monoclonal antibodies of anti-α-catenin, anti-ß-catenin, anti-ZO-1, anti-N-cadherin, anti-JUP, and anti-Cx43 (Invitrogen, San Francisco, CA, USA), rabbit anti-Nav1.5 antibody (Abcam, MA, USA) and mouse anti-GAPDH (SIGMA, Saint Louis, Missouri, USA) for detection of these proteins in immunoassays.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!