Flow (Intracellular), Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Simple Western, Western Blot

Cited:

Flow Cytometry, IF/IHC, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ Clone # 13A9

Concentration

1 mg/ml

View all N-Cadherin Antibodies »

Product Specifications

Immunogen

This N-Cadherin Antibody (13A9) was developed against the cytoplasmic domain of human N Cadherin [Swiss-Prot# P19022].

Localization

Cell membrane; Single-pass type I membrane protein.

Marker

Mesenchymal Cells Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Theoretical MW

140 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.

Scientific Data Images for N-Cadherin Antibody (13A9)

Immunocytochemistry: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunocytochemistry: N-Cadherin Antibody (13A9) [NBP1-48309] - Immunostaining of original tumor, low passage, and high passage KCI-MENG1 cells. The original patient-derived tumor (top row) showed moderate and patchy immunoreactivity for epithelial membrane antigen (EMA); strong and diffuse immunostaining for progesterone receptor (PR); and a Ki-67 proliferative index of 2-3%. There was also strong immunostaining for N-cadherin and vimentin. KCI-MENG1-LP cells (middle row) and KCI-MENG1-HP cells (bottom row) maintained expression of EMA, N-cadherin, and vimentin but had significantly reduced PR expression compared to the original tumor. Whereas Ki-67 labeling was found in only a small number of cells in the original tumor and low passage cells, it was positive in virtually all P84 cells. Scale bar 50 um. Image collected and cropped by CiteAb from the following publication (https://www.translational-medicine.com/content/13/1/227), licensed under a CC-BY license.

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309]

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309] - Western blots showing a reduction in epithelial mesenchymal transition (EMT) markers upon PDK1 knockdown in Colo205 and HT29 cells using E cadherin (NBP2-19051), N cadherin (NBP1-48309) and B-actin antibody (NB600-501). The corresponding secondary antibodies used were either goat anti-rabbit IgG-HRP (NB7160) or goat anti-mouse IgG-HRP (NB7539). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242) licensed under a CC-BY license.

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309]

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309] - Analysis of N-Cadherin expression in HeLa whole cell lysate.

Immunocytochemistry: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunocytochemistry: N-Cadherin Antibody (13A9) [NBP1-48309] - Human meningioma mouse xenograft model KCI-MENG1-LPSX generated with the spontaneously immortal cell line KCI-MENG1-LP. Tumors from immunocompromised SCID mice were dissected and the derivative cell line KCI-MENG1-LPSX CL was generated. The EMA, PR, and N-cadherin IHC of the mouse tumor highly resembled the original patient-derived tumor. The vimentin- and Ki-67-stained cells in the mouse tumor tissue were markedly more abundant and more intensely stained than in the original tumor. KCI-MENG1-LPSX CL cells displayed the same patterns of immunostaining as the high passage parent cell line KCI-MENG1-HP, including the loss of PR staining. Scale bar 50 um. Image collected and cropped by CiteAb from the following publication (https://www.translational-medicine.com/content/13/1/227), licensed under a CC-BY license.

Flow (Intracellular): N-Cadherin Antibody (13A9) [NBP1-48309]

Flow (Intracellular): N-Cadherin Antibody (13A9) [NBP1-48309] - An intracellular stain was performed on HeLa with NBP1-48309 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody.

Simple Western: N-Cadherin Antibody (13A9) [NBP1-48309]

Simple Western: N-Cadherin Antibody (13A9) [NBP1-48309] - Simple Western lane view shows a specific band for N Cadherin in 1.0 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309]

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309] - Mitochondrial fission regulates migration and invasion in BRAF^V600E CRC cells through glucose metabolic reprograming. Western blot showing a reduction in EMT markers upon vemurafenib treatment in BRAF^V600E cells. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33738242/) licensed under a CC-BY license.

Immunocytochemistry/ Immunofluorescence: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunocytochemistry/Immunofluorescence: N-Cadherin Antibody (13A9) [NBP1-48309] - Adult mouse neural stem cells stained for N-Cadherin. Antibody at 1:100. ICC/IF image submitted by a verified customer review.

Immunohistochemistry: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunohistochemistry: N-Cadherin Antibody (13A9) [NBP1-48309] - Immunohistochemical characterization of tissue cultures. Nuclei labeled with DAPI (blue). Whole-mount immunostaining of a cardiomyocyte-only spheroid 1 month in culture, N-cadherin (red) and EH-myomesin (green). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32118040/) licensed under a CC-BY license.

Flow Cytometry: N-Cadherin Antibody (13A9) [NBP1-48309]

Flow Cytometry: N-Cadherin Antibody (13A9) [NBP1-48309] - An intracellular stain was performed on U-251 MG cells with N-Cadherin Antibody (13A9) NBP1-48309AF594 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 594.

Immunocytochemistry/ Immunofluorescence: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunocytochemistry/Immunofluorescence: N-Cadherin Antibody (13A9) [NBP1-48309] - HEK 293 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with anti- at 5 ug/mL overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunohistochemistry: N-Cadherin Antibody (13A9) [NBP1-48309] - IHC analysis of N Cadherin in mouse liver using DAB with hematoxylin counterstain.

Immunohistochemistry-Paraffin: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunohistochemistry-Paraffin: N-Cadherin Antibody (13A9) [NBP1-48309] - Analysis of a FFPE tissue section of human brain using 1:200 dilution of N-Cadherin antibody. The staining was developed using HRP labeled anti-mouse secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.

Flow Cytometry: N-Cadherin Antibody (13A9) [NBP1-48309]

Flow Cytometry: N-Cadherin Antibody (13A9) [NBP1-48309] - An intracellular stain was performed on HeLa cells with NBP1-48309AF488 and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

View 11 more images

Applications for N-Cadherin Antibody (13A9)

Application

Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:50-1:200

Immunohistochemistry-Paraffin

1:50-1:100

Immunoprecipitation

1:10-1:500

Simple Western

1:50

Western Blot

0.5 ug/ml

Application Notes

In Western Blot a band is observed at approx. 140 kDa.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.

Reviewed Applications

Read 2 reviews rated 5 using NBP1-48309 in the following applications:

Immunocytochemistry (1 Review)
Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: N-Cadherin

N-Cadherin, also referred to as Neural Cadherin (NCAD) or Cadherin-2 (CHD2), is a 130 kDa protein that is a member of the calcium-dependent adhesion molecule family of classical (type I) cadherins (1-4). Under the CDH2 gene, human N-cadherin is synthesized as a 906 amino acid protein with a theoretical molecular weight of 99.8 kDa (5). The N-cadherin protein structure is similar to other classical type I cadherins including epithelial (E-) cadherin and placental (P-) cadherin (1,2). N-cadherin consists of a 25 amino acid (aa) N-terminal signal peptide and 134 aa pro-peptide, a 565 aa extracellular domain (ECD) with five cadherin repeats, a 21 aa transmembrane segment, and a 161 aa cytoplasmic domain (1-3,5). The ECD of N-cadherin monomers is responsible for homotypic binding through either cis or trans adhesion (2,3).

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

Entrez Gene IDs

12558 (Mouse)

Gene Symbol

CDH2

UniProt

P19022

Additional N-Cadherin Products

Product Documents for N-Cadherin Antibody (13A9)

Product Datasheets

Download Product Datasheets

Protocols & Manuals

Download NBP1-48309 Specific Protocol

COA

SDS

Download SDS

Product Specific Notices for N-Cadherin Antibody (13A9)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations
Reviews

N-Cadherin Antibody (13A9)

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-48309

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

Concentration

View all N-Cadherin Antibodies »

Product Specifications

Immunogen

Localization

Marker

Clonality

Host

Isotype

Theoretical MW

Scientific Data Images for N-Cadherin Antibody (13A9)

Immunocytochemistry: N-Cadherin Antibody (13A9) [NBP1-48309]

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309]

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunocytochemistry: N-Cadherin Antibody (13A9) [NBP1-48309]

Flow (Intracellular): N-Cadherin Antibody (13A9) [NBP1-48309]

Simple Western: N-Cadherin Antibody (13A9) [NBP1-48309]

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunocytochemistry/ Immunofluorescence: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunohistochemistry: N-Cadherin Antibody (13A9) [NBP1-48309]

Flow Cytometry: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunocytochemistry/ Immunofluorescence: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunohistochemistry: N-Cadherin Antibody (13A9) [NBP1-48309]

Immunohistochemistry-Paraffin: N-Cadherin Antibody (13A9) [NBP1-48309]

Flow Cytometry: N-Cadherin Antibody (13A9) [NBP1-48309]

Applications for N-Cadherin Antibody (13A9)

Immunocytochemistry/ Immunofluorescence

Immunohistochemistry

Immunohistochemistry-Paraffin

Immunoprecipitation

Simple Western

Western Blot

Reviewed Applications

Formulation, Preparation, and Storage

Purification

Formulation

Preservative

Concentration

Shipping

Stability & Storage

Background: N-Cadherin

Long Name

Alternate Names

Entrez Gene IDs

Gene Symbol

UniProt

Additional N-Cadherin Products

Product Documents for N-Cadherin Antibody (13A9)

Product Datasheets

Protocols & Manuals

COA

Certificate of Analysis

SDS

Product Specific Notices for N-Cadherin Antibody (13A9)