Invitrogen

N-cadherin Monoclonal Antibody (8C11)

Product Image Gallery

N-cadherin Antibody in Immunofluorescence (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) — Western blot analysis was performed on tissue lysate and whole cell extract (30 µg) of Mouse Brain (Lane 1) and MCF-7 (Lane 2). The blots were probed with N-cadherin/CDH2 Mouse monoclonal Antibody (Product # MA1-2002, 1 in 1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A ~ 130 kDa band corresponding to N-cadherin/CDH2 was observed across all tissues and cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by overnight transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

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).

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

1:50-1:200

Immunofluorescence (IF)

1:50 - 1:200

Immunohistochemistry (Paraffin) (IHC (P))

1:20

Immunoprecipitation (IP)

5 µg

Western Blot (WB)

1:1000

Published Applications

Immunohistochemistry (IHC)

See 1 publication below

Western Blot (WB)

See 2 publications below

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.

Bioinformatics

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

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

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

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

Molecular Function: cadherin