Invitrogen

N-cadherin Monoclonal Antibody (13A9)

Advanced Verification

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

2 Published Figures

4 References

View all (57) N-cadherin antibodies

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Datasheet

Protocols

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Tech Support

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N-cadherin Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of N-cadherin was performed using 90% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with N-cadherin Mouse Monoclonal Antibody (13A9) (Product # MA1-159) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

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 SH-SY5Y Cas9 (Lane 1) andSH-SY5Y N-Cadherin KO (Lane 2) 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 (13A9) (Product # MA1-159, 1:250 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 on HeLa, MCF7, and NIH/3T3 cells. Antigen-antibody complexes were formed by incubating 300 µg of HeLa, MCF7, and NIH/3T3 whole cell lysates with 2 µg of an N-cadherin monoclonal antibody (Product # MA1-159) overnight on a rocking platform at 4C. 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). Eluted samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with StartingBlock T20 (Product # 37543) for 1 hour. The membrane was probed with an N-cadherin monoclonal antibody (Product # MA1-159) at a concentration of 2 µg/mL overnight rotating at 4C, washed in TBST, and probed with an HRP-conjugated anti-mouse light chain antibody at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

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 (13A9) (Product # MA1-159). {KO}

Product Details

MA1-159

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

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human
Host/Isotype	Mouse / IgG1
Class	Monoclonal
Type	Antibody
Clone	13A9
Immunogen	MBP fusion protein containing the full cytoplasmic domain of human N-cadherin View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein A
Storage buffer	PBS with 1mg/mL BSA, 30% glycerol
Contains	0.05% sodium azide
Storage conditions	-20°C
RRID	AB_2536867

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

Mechano-signalling, induced by fullerene C₆₀ nanofilms, arrests the cell cycle in the G2/M phase and decreases proliferation of liver cancer cells.

International journal of nanomedicine

Sosnowska M,Kutwin M,Jaworski S,Strojny B,Wierzbicki M,Szczepaniak J,Łojkowski M,Święszkowski W,Bałaban J,Chwalibog A,Sawosz E

Published figure using N-cadherin monoclonal antibody (Product # MA1-159) in Western Blot

Fri Nov 08 00:00:00 EST 2019

Diamond, graphite, and graphene oxide nanoparticles decrease migration and invasiveness in glioblastoma cell lines by impairing extracellular adhesion.

International journal of nanomedicine

Wierzbicki M,Jaworski S,Kutwin M,Grodzik M,Strojny B,Kurantowicz N,Zdunek K,Chodun R,Chwalibog A,Sawosz E

Published figure using N-cadherin monoclonal antibody (Product # MA1-159) in Western Blot

Fri Jan 26 00:00:00 EST 2018

Effect of N-cadherin misexpression by the mammary epithelium in mice.

Journal of cellular biochemistry

Knudsen KA,Sauer C,Johnson KR,Wheelock MJ

MA1-159 was used in immunohistochemistry - paraffin section to investigate the effect of N-cadherin misexpression in mice

Mon Aug 15 00:00:00 EDT 2005

P-cadherin expression in breast carcinoma indicates poor survival.

Cancer

Peralta Soler A,Knudsen KA,Salazar H,Han AC,Keshgegian AA

MA1-159 was used in immunohistochemistry - paraffin section to discuss the role of cadherins in tumor development.

Fri Oct 01 00:00:00 EDT 1999

Bioinformatics

Protein Aliases: 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; N-cadherin; NCAD

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

Entrez Gene ID: (Human) 1000, (Mouse) 12558

Molecular Function: cadherin