After harvest, cells were lysed in 20 mmol/L Tris-HCl (pH 7.5) plus protease inhibitor (Solarbio). The extract was centrifuged at 4° for 20 min at 16,000 g. Protein concentration in the supernatant was determined by bicinchoninic acid (BCA) method. The samples were separated by SDS-PAGE. Afterwards, the separated protein was then transferred onto nitrocellulose (NC) membrane. Non-specific binding sites were blocked with 5% skim milk. The membrane was probed with primary and secondary antibodies and then visualized with enhanced chemiluminescence (ECL) reagent. β-actin was used as the internal reference, and the relative expression level of the target protein was presented as gray value of test band/gray value of the β-actin band. Primary antibodies of HMGB3 (ab18256), Caspase 3 (ab32351), Caspase 9 (ab138412), E-cadherin (ab231303), B-cell lymphoma-2 (Bcl-2) (ab182858), BCL2-Associated X (Bax) (ab32503), N-cadherin (ab76011), β-catenin (ab32572), c-myc (ab32072), Matrix metalloproteinase 7 (MMP7) (ab207299), β-actin (ab8226), and secondary antibody (HRP conjugate) were all purchased from Abcam.
N cadherin ab76011
Manufactured by Abcam
Sourced in United Kingdom
N-cadherin (ab76011) is a primary antibody that recognizes the N-cadherin protein. N-cadherin is a type-I classical cadherin that plays a role in cell-cell adhesion.
Automatically generated - may contain errors
Lab products found in correlation
Ripa lysis buffer, by Beyotime (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ab207299, by Abcam (1 mentions)
Ab32351, by Abcam (1 mentions)
Ab182858, by Abcam (1 mentions)
Ab18256, by Abcam (1 mentions)
Ab32503, by Abcam (1 mentions)
Myc ab32072, by Abcam (1 mentions)
β catenin ab32572, by Abcam (1 mentions)
Protease inhibitor, by Solarbio (1 mentions)
Ab28484, by Abcam (1 mentions)
Vimentin ab92547, by Abcam (1 mentions)
E cadherin ab40772, by Abcam (1 mentions)
Ripa lysis buffer, by Solarbio (1 mentions)
β actin antibody, by Cell Signaling Technology (1 mentions)
E cadherin ab1416, by Abcam (1 mentions)
Ecl developer, by Thermo Fisher Scientific (1 mentions)
Odyssey gel imaging system, by Thermo Fisher Scientific (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Ab128915, by Abcam (1 mentions)
4 protocols using n cadherin ab76011
1
Western Blot Analysis of Cellular Proteins
2
Protein Expression Analysis of Cartilage Extracellular Matrix
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in RIPA lysis buffer (Solarbio, Beijing) and centrifuged to extract total protein. The protein concentration of the samples was determined using the BCA method. Protein samples of the same quality were separated in 8% SDS-PAGE and transferred to PVDF membranes. Membranes were blocked with 2% nonfat dry milk in TBST, incubated with primary antibodies overnight at 4°C, and secondary antibodies were added the next day for 1 h at room temperature. The bands were photographed under appropriate conditions using a luminescent reagent (Solarbio, Beijing). Antibodies to detect PRG4 (ab28484), E-cadherin (ab40772), N-cadherin (ab76011), vimentin (ab92547), and β-actin (ab8226) were purchased from Abcam (Shanghai).
3
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total exosomic and cellular protein was extracted using RIPA lysis buffer (Beyotime Institute of Biotechnology, Beijing, China). The protein concentrations of cell and exosome lysates were determined using BCA protein assay kit (Beyotime Institute of Biotechnology, Beijing, China). For Western blot analyses, the cellular and exosomic proteins (10 µg/well) were separated by electrophoresis in 10% sodium dodecylsulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membrane (Amersham, Buckinghamshire, UK). The membrane was blocked with 5% skimmed milk in TBS-T (10 mM TrisCl, pH 8.0, 150 mM NaCl, 0.5% Tween 20) at 4°C overnight, rinsed 3 times (10 min/time) with TBS-T, followed by 3 h incubation at room temperature with the specific primary antibodies, followed by 1 h incubation with the second antibody. ECL developer (Thermo Scientific) was added and exposure imaging was performed by Odyssey gel imaging system. The specific antibodies to E-cadherin (ab1416), N-cadherin (ab76011), Vimentin (ab92547), Snail (ab216347), CD81 (ab109201), HSP70 (ab181606), TSG101 (ab125011), Calnexin (ab133615), were purchased from Abcam (Cambridge, UK). β-actin antibody (#4970) was acquired from Cell Signaling Technology (Danvers, MA, USA).
4
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were extracted using RIPA lysis buffer (Beyotime Biotechnology, China) supplemented with protease inhibitors (Roche, China). Then the collected proteins were quantified with a BCA™ Protein Assay Kit (#23227, Thermo Fisher, USA). Proteins were separated by 12% sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE), and then transferred onto the polyvinylidene difluoride (PVDF) membranes. After blocking with skim milk, membranes were incubated with the primary antibodies overnight at 4 C. The primary antibodies to E-cadherin (ab40772), N-cadherin (ab76011) and GAPDH (ab128915) were all purchased from Abcam Company (Abcam, Cambridge, UK). After that, these membranes were incubated with secondary antibody for 2 h at room temperature. The signals were visualized by chemiluminescent detection system (BB-3501; Amersham Pharmacia Biotech, UK). Internal control was GADPH.
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!