Invitrogen

N-cadherin Monoclonal Antibody (13A9)

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

1 Published Figure

3 References

View all (47) N-cadherin antibodies

Datasheet

Datasheet

Product Image Gallery

N-cadherin Antibody — Figure 5 EGFR/AKT/mTOR and beta-catenin signaling in glioblastoma cells after treated with nanoparticles. Notes: ( A ) Confocal microscope images of U87 and U118 cells actin cytoskeleton. Cells were grown on extracellular matrix for 24 h and treated with diamond nanoparticles, graphite nanoparticles, or graphene oxide nanoparticles at a concentration of 20 mug/mL and incubated for 24 h. F-Actin was stained with phalloidin conjugated with Atto 633. ( B ) Western blot analysis of N-cadherin, pan-cadherin, vinculin, p-EGFR, and EGFR. GAPDH was used as a loading control. ( C ) Western blot analysis of nuclear and cytoplasmic protein fractions used for determination of beta-catenin protein level. PCNA and beta-tubulin were used as loading controls for nuclear and cytoplasmic fractions, respectively. ( D ) ELISA analysis of AKT and mTOR phosphorylation in comparison to control. Treatment with nanoparticles significantly reduced phospho-AKT ( P <0.0000) and mTOR ( P <0.0000). Phospho-AKT level was cell line dependent ( P <0.0000) and was affected by interaction of cell line and nanoparticles ( P <0.001). Phospho-mTOR level was also cell line dependent ( P <0.0023) and was affected by interaction of cell line and nanoparticles ( P <0.0000). Values are expressed as mean +- standard deviation (n=5, each experiment in duplicate). Statistical significance is indicated with different superscripts (multifactor ANOVA; P <0.05). Abbreviations: ANOVA, analysis of variance; C, control; ELISA,

N-cadherin Antibody in Immunofluorescence (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 # MA5-159) 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) — Western blot analysis of N-cadherin was performed by loading 25 µg of the indicated tissue lysate or whole cell lysates and 15 µL of PageRuler Prestained Protein Ladder (Product # 26616) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 1 hour. The membrane was probed with an N-cadherin monoclonal antibody (Product # MA1-159) at a concentration of 3 µg/mL overnight at 4C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

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 in Independent antibody — Antibody specificity was demonstrated by showing that antibodies raised against the same target protein perform similarly. Western blot of N-cadherin using N-cadherin Monoclonal Antibody (Product # MA1-159), tested in parallel with N-cadherin Monoclonal Antibody (Product # MA5-15633), shows similar expression of N-cadherin in the tissue tested. {IAV}

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

1:50-1:200

Immunofluorescence (IF)

1:50-1:200

Immunoprecipitation (IP)

2 µg

Western Blot (WB)

1:200-1:1000

Published Applications

Western Blot (WB)

See 1 publication below

Immunohistochemistry (Paraffin) (IHC (P))

See 2 publications below

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

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.

Bioinformatics

Protein Aliases: cadherin 2, type 1, N-cadherin (neuronal); Cadherin-2; Calcium-Dependent Adhesion Protein; calcium-dependent adhesion protein, neuronal; CD325; CD325 Neural cadherin; CDw325; 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