Anti-N-Cadherin, clone 13A9

Species Reactivity	Key Applications	Host	Format	Antibody Type
H	ICC, IHC, IP, WB	M	Culture Supernatant	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

ICC, IHC, IP, WB

Culture Supernatant

Monoclonal Antibody

Description
Catalogue Number	05-915
Replaces	04-1126
Brand Family	Upstate
Trade Name	Upstate
Description	Anti-N-Cadherin Antibody, clone 13A9
Alternate Names	Cadherin-2 CD325 CDw325 N-cadherin Neural cadherin
Background Information	Cadherin-2 (UniProt P19022; also known as CD325, CDw325, N-cadherin, Neural cadherin) is encoded by the CDH2 (also known as CDHN, NCAD) gene (Gene ID 1000) in human. Cadherins constitute a family of calcium-dependent cell-cell adhesion proteins that play important roles in the embryonic development and maintenance of normal tissue architecture. Cadherins are composed of an extracellular domain (a.a. 160-724 of human N-cadherin) with five homologous repeats that mediates adhesion, a single pass transmembrane domain (a.a. 725-745 of human N-cadherin), and a conserved cytoplasmic domain (a.a. 746-906 of human N-cadherin) that interacts with catenins to link cadherins to the actin cytoskeleton. In addition, a known Src substrate p120ctn also modulate the strength of cadherin-dependent adhesion by interacting with cadherins at their intracellular juxtamembrane domain. Cadherins are synthesized as precursor proteins that must be proteolytically cleaved to generate functional, mature proteins. Newly synthesized proN-cadherin (a.a. 1-906) is phosphorylated and proteolytically processed prior to transport to the plasma membrane. In addition, Plakoglobin (gamma-catenin) and beta-catenin associate only with phosphorylated proN-cadherin, whereas p120ctn can associate with both phosphorylated and non-phosphorylated proN-cadherin. The N-terminal signal and propeptide (a.a. 1-25 and 26-159 of of human N-cadherin) region is proteolytically removed and a core N-cadherin-catenin complex is assembled in the endoplasmic reticulum or Golgi compartment prior to localization at the plasma membrane where linkage to the actin cytoskeleton can be established.

Description

Catalogue Number

05-915

Replaces

04-1126

Brand Family

Upstate

Trade Name

Upstate

Description

Anti-N-Cadherin Antibody, clone 13A9

Alternate Names

Cadherin-2
CD325
CDw325
N-cadherin
Neural cadherin

Background Information

Cadherin-2 (UniProt P19022; also known as CD325, CDw325, N-cadherin, Neural cadherin) is encoded by the CDH2 (also known as CDHN, NCAD) gene (Gene ID 1000) in human. Cadherins constitute a family of calcium-dependent cell-cell adhesion proteins that play important roles in the embryonic development and maintenance of normal tissue architecture. Cadherins are composed of an extracellular domain (a.a. 160-724 of human N-cadherin) with five homologous repeats that mediates adhesion, a single pass transmembrane domain (a.a. 725-745 of human N-cadherin), and a conserved cytoplasmic domain (a.a. 746-906 of human N-cadherin) that interacts with catenins to link cadherins to the actin cytoskeleton. In addition, a known Src substrate p120ctn also modulate the strength of cadherin-dependent adhesion by interacting with cadherins at their intracellular juxtamembrane domain. Cadherins are synthesized as precursor proteins that must be proteolytically cleaved to generate functional, mature proteins. Newly synthesized proN-cadherin (a.a. 1-906) is phosphorylated and proteolytically processed prior to transport to the plasma membrane. In addition, Plakoglobin (gamma-catenin) and beta-catenin associate only with phosphorylated proN-cadherin, whereas p120ctn can associate with both phosphorylated and non-phosphorylated proN-cadherin. The N-terminal signal and propeptide (a.a. 1-25 and 26-159 of of human N-cadherin) region is proteolytically removed and a core N-cadherin-catenin complex is assembled in the endoplasmic reticulum or Golgi compartment prior to localization at the plasma membrane where linkage to the actin cytoskeleton can be established.

Product Information
Format	Culture Supernatant
Control	HeLa cell lysate
Presentation	Mouse monoclonal immunoglobulin hybridoma culture supernatant containing 0.05% sodium azide before the addition of glycerol to 30%.

Product Information

Format

Culture Supernatant

Control

HeLa cell lysate

Presentation

Mouse monoclonal immunoglobulin hybridoma culture supernatant containing 0.05% sodium azide before the addition of glycerol to 30%.

Applications
Application	Detect N-cadherin using this Anti-N-Cadherin Antibody, clone 13A9 validated for use in Immunocytochemistry, Immunohistochemistry, Immunoprecipitation, and Western Blotting.
Key Applications	Immunocytochemistry Immunohistochemistry Immunoprecipitation Western Blotting
Application Notes	Immunohistochemistry Analysis: A representative lot immunostained the extracellular matrix of the stable plaques and in the fibrous cap region rich in vascular smooth muscle cells (VSMCs) using patients-derived parafﬁn-embedded internal carotid artery tissue sections (Musumeci, G., et al. (2014). Histol. Histopathol. 29(6):707-719). Immunohistochemistry Analysis: A representative lot detected strong N-cadherin immunoreactivity in parafﬁn-embedded rectal cancer (RC) tissues with positive regional lymph node metastasis (RLNM) status, while only weak N-cadherin immunoreactivity was detected in RC with negative RLNM, and no N-cadherin staining was seen in normal colorectal epithelium (Fan, X.J., et al. (2012). Br. J. Cancer. 106(11):1735-1741). Immunohistochemistry Analysis: A representative lot detected N-cadherin immunoreactivity in formalin-ﬁxed, parafﬁn-embedded hepatocellular carcinoma (HCC) tissue sections. A signiﬁcant inverse correlation was found between RUNX3 and N-cadherin expression levels (Tanaka, S., et al. (2012). Int. J. Cancer. 131(11):2537-2546). Western Blotting Analysis: A representative lot detected an upregulated N-cadherin expression in CCL185 carcinoma cells following transient Epstein-Barr virus (EBV) infection. The EMT-like phenotype remained even after viral loss by culture selection pressure withdrawal (Queen, K.J., et al. (2013). Int. J. Cancer. 132(9):2076-2086). Western Blotting Analysis: A representative lot detected N-cadherin in Hep3B, Huh7, HLF and SK-Hep1 human hepatocellular carcinoma (HCC) cell lysates (Tanaka, S., et al. (2012). Int. J. Cancer. 131(11):2537-2546). Western Blotting Analysis: A representative lot detected both the unprocessed (pro-) and processed (mature) forms of N-cadherin in HeLa cell lysate (Wahl, J.K. 3rd., et al. (2003). J. Biol. Chem. 278(19):17269-17276). Western Blotting Analysis: A representative lot detected N-cadherin in WI-38 human fibroblast lysate, but not in JAr human placental choriocarcinoma cell lysate (Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77). Immunocytochemistry Analysis: A representative lot detected N-cadherin immunoreactivity localized primarily at the cell-cell borders by fluorescent immunocytochemistry staining of 1% paraformaldehyde-fixed, methanol-permeabilized HeLa cells (Wahl, J.K. 3rd., et al. (2003). J. Biol. Chem. 278(19):17269-17276). Immunocytochemistry Analysis: A representative lot detected N-cadherin immunoreactivity colocalized with those of alpha- and beta-catenin by dual fluorescent immunocytochemistry staining of fixed WI-38 human fibroblasts (Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77). Immunoprecipitation Analysis: Representative lots co-immunoprecipitated alpha-catenin, beta-catenin, and plakoglobin with N-cadherin from WI-38 human fibroblast and HeLa cell lysates (Wahl, J.K. 3rd., et al. (2003). J. Biol. Chem. 278(19):17269-17276; Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77).

Applications

Application

Detect N-cadherin using this Anti-N-Cadherin Antibody, clone 13A9 validated for use in Immunocytochemistry, Immunohistochemistry, Immunoprecipitation, and Western Blotting.

Key Applications

Immunocytochemistry
Immunohistochemistry
Immunoprecipitation
Western Blotting

Application Notes

Immunohistochemistry Analysis: A representative lot immunostained the extracellular matrix of the stable plaques and in the fibrous cap region rich in vascular smooth muscle cells (VSMCs) using patients-derived parafﬁn-embedded internal carotid artery tissue sections (Musumeci, G., et al. (2014). Histol. Histopathol. 29(6):707-719).
Immunohistochemistry Analysis: A representative lot detected strong N-cadherin immunoreactivity in parafﬁn-embedded rectal cancer (RC) tissues with positive regional lymph node metastasis (RLNM) status, while only weak N-cadherin immunoreactivity was detected in RC with negative RLNM, and no N-cadherin staining was seen in normal colorectal epithelium (Fan, X.J., et al. (2012). Br. J. Cancer. 106(11):1735-1741).
Immunohistochemistry Analysis: A representative lot detected N-cadherin immunoreactivity in formalin-ﬁxed, parafﬁn-embedded hepatocellular carcinoma (HCC) tissue sections. A signiﬁcant inverse correlation was found between RUNX3 and N-cadherin expression levels (Tanaka, S., et al. (2012). Int. J. Cancer. 131(11):2537-2546).
Western Blotting Analysis: A representative lot detected an upregulated N-cadherin expression in CCL185 carcinoma cells following transient Epstein-Barr virus (EBV) infection. The EMT-like phenotype remained even after viral loss by culture selection pressure withdrawal (Queen, K.J., et al. (2013). Int. J. Cancer. 132(9):2076-2086).
Western Blotting Analysis: A representative lot detected N-cadherin in Hep3B, Huh7, HLF and SK-Hep1 human hepatocellular carcinoma (HCC) cell lysates (Tanaka, S., et al. (2012). Int. J. Cancer. 131(11):2537-2546).
Western Blotting Analysis: A representative lot detected both the unprocessed (pro-) and processed (mature) forms of N-cadherin in HeLa cell lysate (Wahl, J.K. 3rd., et al. (2003). J. Biol. Chem. 278(19):17269-17276).
Western Blotting Analysis: A representative lot detected N-cadherin in WI-38 human fibroblast lysate, but not in JAr human placental choriocarcinoma cell lysate (Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77).
Immunocytochemistry Analysis: A representative lot detected N-cadherin immunoreactivity localized primarily at the cell-cell borders by fluorescent immunocytochemistry staining of 1% paraformaldehyde-fixed, methanol-permeabilized HeLa cells (Wahl, J.K. 3rd., et al. (2003). J. Biol. Chem. 278(19):17269-17276).
Immunocytochemistry Analysis: A representative lot detected N-cadherin immunoreactivity colocalized with those of alpha- and beta-catenin by dual fluorescent immunocytochemistry staining of fixed WI-38 human fibroblasts (Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77).
Immunoprecipitation Analysis: Representative lots co-immunoprecipitated alpha-catenin, beta-catenin, and plakoglobin with N-cadherin from WI-38 human fibroblast and HeLa cell lysates (Wahl, J.K. 3rd., et al. (2003). J. Biol. Chem. 278(19):17269-17276; Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77).

Biological Information
Immunogen	Bacterially expressed human N-cadherin cytoplasmic domain MBP fusion protein (Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77).
Epitope	Cytoplasmic domain.
Clone	13A9
Host	Mouse
Specificity	Clone 13A9 recognizes N-cadherin, but not P-, E-, or M-cadherin (Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77).
Species Reactivity	Human
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NM_001792
Gene Symbol	CDH2 CDHN NCAD
Purification Method	Unpurified
UniProt Number	P19022
Molecular Weight	~140 kDa observed. Target band size appears larger than the calculated molecular weights of 82.03 kDa (mature) and 99.81/97.04 kDa (isoform 1/2 pro-form) due to posttranslational glycosylation and phosphorylation.

Biological Information

Immunogen

Bacterially expressed human N-cadherin cytoplasmic domain MBP fusion protein (Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77).

Epitope

Cytoplasmic domain.

Clone

13A9

Host

Mouse

Specificity

Clone 13A9 recognizes N-cadherin, but not P-, E-, or M-cadherin (Knudsen, K.A., et al. (1995). J. Cell Biol. 130(1):67-77).

Species Reactivity

Human

Antibody Type

Monoclonal Antibody

Entrez Gene Number

NM_001792

Gene Symbol

CDH2
CDHN
NCAD

Purification Method

Unpurified

UniProt Number

P19022

Molecular Weight

~140 kDa observed. Target band size appears larger than the calculated molecular weights of 82.03 kDa (mature) and 99.81/97.04 kDa (isoform 1/2 pro-form) due to posttranslational glycosylation and phosphorylation.

Product Usage Statements
Quality Assurance	Evaluated by Western Blotting in HeLa cell lysate. Western Blotting Analysis: A 1:1000-5000 dilution of this hybridoma culture supernatant detected N-cadherin in HeLa cell lysate.
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Quality Assurance

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: A 1:1000-5000 dilution of this hybridoma culture supernatant detected N-cadherin in HeLa cell lysate.

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Maintain for 2 years at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Storage and Shipping Information

Storage Conditions

Maintain for 2 years at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Packaging Information
Material Size	100 µL

Packaging Information

Material Size

100 µL

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

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Anti-N-Cadherin, clone 13A9

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