Invitrogen

N-cadherin Monoclonal Antibody (8C11)

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

1 Published Figure

6 References

View all (57) N-cadherin antibodies

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N-cadherin Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of N-Cadherin in SHSY5Y cells (human neuroblast). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) an N-Cadherin monoclonal antibody (Product # MA1-2002) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.

N-cadherin Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of N-cadherin showing staining in the membrane of paraffin-embedded human heart tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a N-cadherin Mouse Monoclonal Antibody (Product # MA1-2002) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

N-cadherin Antibody in Western Blot (WB) — Knockout of N-cadherin was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR914282_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of N-cadherin was performed by loading 30 µg of SHSY5Y Wild Type (Lane 1), SHSY5Y Cas9 (Lane 2) andSHSY5Y N-Cadherin KO (Lane 3) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-N-cadherin Monoclonal Antibody (8C11) (Product # MA1-2002, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to N-cadherin.

N-cadherin Antibody in Immunoprecipitation (IP) — Immunoprecipitation of N-Cadherin was performed using SHSY5Y whole cell lysate. Antigen-antibody complexes were formed by incubating 300 µg of lysate with 5 µg of an N-Cadherin monoclonal antibody (Product # MA1-2002) overnight on a rocking platform at 4°C. The immune complexes were captured on 50 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). The sample was resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with an N-cadherin monoclonal antibody (Product # MA1-2002) at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with Clean-blot IP Detection Reagent (Product # 21230) at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).

N-cadherin Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein inN-cadherin KO cell line compared to control cell line using Anti-N-cadherin Monoclonal Antibody (8C11) (Product # MA1-2002). {KO}

Product Details

MA1-2002

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	View 4 publications 4 publications
Immunohistochemistry (IHC)	-	View 2 publications 2 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:20	-
Immunocytochemistry (ICC/IF)	1:50-1:200	-
Immunoprecipitation (IP)	5 µg	-

Product Specifications
Species Reactivity	Hamster, Human, Mouse, Rabbit
Published species	Avian, Human
Host/Isotype	Mouse / IgG1
Class	Monoclonal
Type	Antibody
Clone	8C11
Immunogen	Bacterial fusion protein corresponding to the extracellular domain of N-cadherin.
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein G
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_326076

Product Specific Information

MA1-2002 detects N-Cadherin from human neuronal samples, including SHSY5Y cells. MA1-2002 also detects N-Cadherin in rabbit and hamster brain lysates. MA1-2002 does not detect N-Cadherin in lysates from mouse, rat, pig and cow brain. This antibody is not suitable for mouse C2C12 cell lysates in Western blot analysis.

MA1-2002 has been successfully used in Western blot, Immunoprecipitation, IHC (P) and Immunofluorescence applications. By Western blot, this antibody detects N-Cadherin at 130 kDa. This antibody is not suitable for frozen tissues in immunohistochemistry applications.

The MA1-2002 immunogen is a bacterial fusion protein to the extracellular domain of N-cadherin (a domain between EC3 and EC4, amino acid residues 92-593 from human N-cadherin).

Target Information

N-cadherin is a 140 kDa protein belonging to a family of transmembrane molecules that mediate calcium-dependent intercellular adhesion. Cadherins are involved in controlling morphogenetic movements during development and regulate cell surface adhesion through homotypic adhesion with the same cadherin species. N-cadherin's function is dependent on its association with the actin-cytoskeleton and is mediated through interactions between the C-terminal region of N-cadherin and the cytoplasmic catenin proteins. The stability of this association is regulated by phosphorylation and dephosphorylation of beta-catenin. Further, N-cadherin is a calcium dependent cell-cell adhesion glycoprotein comprised of five extracellular cadherin repeats, a transmembrane region and a highly conserved cytoplasmic tail. The protein functions during gastrulation and is required for establishment of left-right asymmetry.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

The Motility and Mesenchymal Features of Breast Cancer Cells Correlate with the Levels and Intracellular Localization of Transglutaminase Type 2.

Cells

Bianchi N,Brugnoli F,Grassilli S,Bourgeois K,Keillor JW,Bergamini CM,Aguiari G,Volinia S,Bertagnolo V

MA1-2002 was used in Western Blot to investigate motility in breast cancer cell lines in association with the expression of Transglutaminase type 2 as well as upon the administration of Doxorubicin, an active cytotoxic agent that is employed in chemotherapy.

Sat Nov 06 00:00:00 EDT 2021

Exosome-mediated mRNA delivery in vivo is safe and can be used to induce SARS-CoV-2 immunity.

The Journal of biological chemistry

Tsai SJ,Atai NA,Cacciottolo M,Nice J,Salehi A,Guo C,Sedgwick A,Kanagavelu S,Gould SJ

MA1-2002 was used in Western Blot to describe an alternative approach using exosomes, the only biologically normal nanovesicle.

Mon Nov 01 00:00:00 EDT 2021

Exosome-mediated mRNA delivery in vivo is safe and can be used to induce SARS-CoV-2 immunity.

The Journal of biological chemistry

Tsai SJ,Atai NA,Cacciottolo M,Nice J,Salehi A,Guo C,Sedgwick A,Kanagavelu S,Gould SJ

MA1-2002 was used in Western Blot to describe an alternative approach using exosomes, the only biologically normal nanovesicle.

Mon Nov 01 00:00:00 EDT 2021

A mechanically active heterotypic E-cadherin/N-cadherin adhesion enables fibroblasts to drive cancer cell invasion.

Nature cell biology

Labernadie A,Kato T,Brugués A,Serra-Picamal X,Derzsi S,Arwert E,Weston A,González-Tarragó V,Elosegui-Artola A,Albertazzi L,Alcaraz J,Roca-Cusachs P,Sahai E,Trepat X

MA1-2002 was used in Immunohistochemistry to investigate the interactions between cancer-associated fibroblasts and cancer cells which drive cooperative tumour invasion.

Wed Mar 01 00:00:00 EST 2017

Control of cell-cell forces and collective cell dynamics by the intercellular adhesome.

Nature cell biology

Bazellières E,Conte V,Elosegui-Artola A,Serra-Picamal X,Bintanel-Morcillo M,Roca-Cusachs P,Muñoz JJ,Sales-Pardo M,Guimerà R,Trepat X

MA1-2002 was used in immunohistochemistry to elucidate the contributions of cell-cell adhesion proteins to intercellular force and epithelial tissue dynamics.

Wed Apr 01 00:00:00 EDT 2015

Distinct PTPmu-associated signaling molecules differentially regulate neurite outgrowth on E-, N-, and R-cadherin.

Molecular and cellular neurosciences

Oblander SA,Brady-Kalnay SM

MA1-2002 was used in western blot to investigate the effects of PTPmu on cadherin-dependent neurite outgrowth

Sat May 01 00:00:00 EDT 2010

N-cadherin-catenin complexes form prior to cleavage of the proregion and transport to the plasma membrane.

The Journal of biological chemistry

Wahl JK,Kim YJ,Cullen JM,Johnson KR,Wheelock MJ

MA1-2002 was used in western blot to investigate N-cadherin processing by proteolysis and the formation of N-cadherin-catenin complexes

Fri May 09 00:00:00 EDT 2003

Bioinformatics

Protein Aliases: cadherin 2, type 1 preproprotein; cadherin 2, type 1, N-cadherin (neuronal); Cadherin-2; Cadherin2; calcium-dependent adhesion protein, neuronal; CD325; CDw325; CDw325 antigen; N-cadherin; N-cadherin 1; Neural cadherin

Gene Aliases: CD325; CDH2; CDHN; CDw325; I79_014914; N-cadherin; NCAD

UniProt ID: (Human) P19022, (Mouse) P15116

Entrez Gene ID: (Human) 1000, (Rabbit) 100344571, (Mouse) 12558, (Chinese hamster) 100689204

Molecular Function: cadherin