N-Cadherin Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-09030
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for N-Cadherin Overexpression Lysate
N-Cadherin Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC207170
Accession#: NM_001792
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Expression Host: HEK293T
Plasmid: RC207170
Accession#: NM_001792
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Product Specifications
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
TMW
97.2 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Type
Overexpression
Scientific Data Images for N-Cadherin Overexpression Lysate
![Western Blot: N-Cadherin Overexpression Lysate [NBL1-09030]](https://resources.bio-techne.com/images/products/N-Cadherin-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-09030-img0002.jpg "Western Blot: N-Cadherin Overexpression Lysate [NBL1-09030]")
Western Blot: N-Cadherin Overexpression Lysate [NBL1-09030]
Western Blot: N Cadherin Overexpression Lysate (Adult Normal) [NBL1-09030] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for N Cadherin.Formulation, Preparation, and Storage
Formulation
RIPA buffer
Concentration
The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store at -80C. Avoid freeze-thaw cycles.
Background: N-Cadherin
N-cadherin is expressed on multiple cell types but is most highly expressed by mesenchymal cells and neural tissue (2). Functionally, N-cadherin has a number of roles including maintaining structural integrity and adhesion, cell signaling, and formation of neuronal synapses and the vascular wall (2). The cytoplasmic tail interacts with beta-catenin which then binds with alpha-catenin, forming the cadherin-catenin adhesion complex, an important component of adhesions junctions (1-3). Given its role in adhesion, N-cadherin serves as an indicator of epithelial-to-mesenchymal transition (EMT) (1-4). The loss of E-cadherin during EMT corresponds with an increase in N-cadherin expression (1-4). This "cadherin-switch" is associated with increased migratory and invasive behavior observed in tumor progress (1-4). Proteases including activity of a disintegrin and metalloprotease 10 (ADAM10), matrix metalloproteinases (MMPs), caspase 3, presenilin, and calpain can cleave N-cadherin as a mechanism for regulating Wnt/beta-catenin signaling and inducing oncogenic signals (3,4). In addition to its expression in solid tumors, N-cadherin has been indicated in hematological disorders such as leukemia and multiple myeloma (1). N-cadherin antagonists are currently being studied as potential therapeutics for a variety of cancer studies (1-2).
References
1. Mrozik, K. M., Blaschuk, O. W., Cheong, C. M., Zannettino, A., & Vandyke, K. (2018). N-cadherin in cancer metastasis, its emerging role in haematological malignancies and potential as a therapeutic target in cancer. BMC Cancer. https://doi.org/10.1186/s12885-018-4845-0
2. Loh, C. Y., Chai, J. Y., Tang, T. F., Wong, W. F., Sethi, G., Shanmugam, M. K., Chong, P. P., & Looi, C. Y. (2019). The E-Cadherin and N-Cadherin Switch in Epithelial-to-Mesenchymal Transition: Signaling, Therapeutic Implications, and Challenges. Cells. https://doi.org/10.3390/cells8101118
3. Derycke, L. D., & Bracke, M. E. (2004). N-cadherin in the spotlight of cell-cell adhesion, differentiation, embryogenesis, invasion and signalling. The International Journal of Developmental Biology. https://doi.org/10.1387/ijdb.041793ld
4. Yu, W., Yang, L., Li, T., & Zhang, Y. (2019). Cadherin Signaling in Cancer: Its Functions and Role as a Therapeutic Target. Frontiers in Oncology. https://doi.org/10.3389/fonc.2019.00989
5. Unitprot (P1903)
Long Name
Neural Cadherin
Alternate Names
Cadherin-2, CD325, CDH2, NCadherin
Gene Symbol
CDH2
Additional N-Cadherin Products
Product Documents for N-Cadherin Overexpression Lysate
Product Specific Notices for N-Cadherin Overexpression Lysate
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.
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