Anti-E-Cadherin (Cytoplasmic) Recombinant Chicken Chimeric mAb (M168)

Name: Anti-E-Cadherin (Cytoplasmic) Recombinant Chicken Chimeric mAb
Brand: Aves Labs
SKU: 78-608-020
Price: 481.0 USD
Availability: InStock

Our chicken Anti-E-Cadherin (Cytoplasmic) recombinant monoclonal antibody is a chimeric antibody derived from clone M168. It detects human, mouse, and rat E-Cadherin (Cytoplasmic) and is purified by affinity chromatography. Works in ELISA, WB, ICC, IHC, IP.

Immunofluorescent staining for detection of cytoskeleton in human colon cancer tissue using Aves Labs anti-E-Cadherin chicken recombinant monoclonal antibody (Cat No. 78-608) at 1:1000 dilution. This section of formalin fixed, paraffin embedded colon cancer tissue was blocked with Immuno Protein Blocking Solution with 5% Goat Serum (AR-6591-04). After washing with TBST, the tissue was stained using goat anti-chicken IgY FITC conjugated secondary antibody (Cat No. 80-1000-FITC) (green). The tissue was mounted with Antibodies Incorporated Fluoroshield with DAPI and DABCO mounting medium (Cat No. AR-6505). DAPI nuclear stain (blue) shows cell nuclei. Magnification 400X.

Immunofluorescent staining of MCF-7 and HeLa cells using Aves Labs anti-E-Cadherin (Cytoplasmic) chicken recombinant antibody [M168](Cat No. 78-608). The cells were fixed with 3% paraformaldehyde, washed with PBS, and blocked/permeabilized with 5% non-fat milk powder in PBST. The anti-E-Cadherin antibody (Cat. 78-608) was incubated with the sample at a dilution of 1:1000 (1 ug/mL) at 4C overnight. After washing with PBST, the cells were stained using goat anti-chicken IgY CF488 secondary antibody (Cat No. 80-1001-488) (green). The cells were mounted with Antibodies Incorporated Fluoroshield with DAPI mounting medium (Cat No. AR-6501). DAPI nuclear stain (blue) shows cell nuclei. The staining revealed the characteristic membranous localization of E-Cadherin in MCF-7 cells, with no staining observed in E-Cadherin negative HeLa cells.

Western blot detection of E-Cadherin in MCF-7 (lane 1), HepG2 (lane 2) and HeLa (lane 3) cell lysates using Aves Labs anti-E-Cadherin (Cytoplasmic) chicken recombinant antibody [M168](Cat No. 78-608). 4-20% Tris-Glycine gel was loaded with 20 ug of MCF-7, HepG2 and HeLa cell lysates. Proteins were transferred to PVDF membrane and blocked with 5% non-fat milk powder. Anti-E-Cadherin chicken antibody was incubated with the membrane at a concentration of 1:1000 (1 ug/ml) and then detected via chemiluminescence (30 seconds) using a goat anti-Chicken H+L, HRP conjugated secondary antibody. The anti-E-Cadherin chicken recombinant antibody specifically detected endogenous E-Cadherin in E-Cadherin-positive MCF-7 and HepG2 cell lysates, but not in E-Cadherin-negative HeLa cell lysates, at the expected molecular weight of ~120 kDa. GAPDH was used as a loading control.

Immunofluorescent staining for detection of cytoskeleton in human colon cancer tissue using Aves Labs anti-E-Cadherin chicken recombinant monoclonal antibody (Cat No. 78-608) at 1:1000 dilution. This section of formalin fixed, paraffin embedded colon cancer tissue was blocked with Immuno Protein Blocking Solution with 5% Goat Serum (AR-6591-04). After washing with TBST, the tissue was stained using goat anti-chicken IgY FITC conjugated secondary antibody (Cat No. 80-1000-FITC) (green). The tissue was mounted with Antibodies Incorporated Fluoroshield with DAPI and DABCO mounting medium (Cat No. AR-6505). DAPI nuclear stain (blue) shows cell nuclei. Magnification 400X.

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Species Reactivity

Human, Mouse, Rat

Applications

ICC, IHC, IP, WB

Host Species

Chicken

Conjugates and Formulations

490

550

594

650

Biotin

Carrier-Free

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Data Sheet

Trial Size

SKU: 78-608-020

Volume: 20 µL

Variant

Price:

$144.00

Quantity:

Ships: 1-2 business days

Product Details

Target

E-Cadherin (Cytoplasmic)

Scientific Background

Cadherins are transmembrane glycoproteins vital in calcium-dependent cell-cell adhesion during tissue differentiation. Cadherins cluster to form foci of homophilic binding units. A key determinant to the strength of the cadherin-mediated adhesion may be by the juxtamembrane region in cadherins. This region induces clustering and also binds to the protein p120 catenin. The cytoplasmic region is highly conserved in sequence and has been shown experimentally to regulate the cell-cell binding function of the extracellular domain of E-cadherin, possibly through interaction with the cytoskeleton. Many cadherins are regulated by phosphorylation, including N-cadherin and E-cadherin. N-cadherin is phosphorylated by c-Src at Tyr-820, Tyr-853, Tyr-860, Tyr-884, and Tyr-886. Phosphorylation of Tyr-860 can disrupt cadherin binding to β-catenin. Since many of these tyrosine sites are conserved in the cadherin family, phosphorylation of these sites may be critical for cadherin function.

Format

Purified by affinity chromatography

Concentration

1 mg/mL

Clonality

Recombinant

Clone

M168

Isotype

IgY

Applications

ICC, IHC, IP, WB

Host Species

Chicken

Gene Name

CDH1

Molecular Weight

120 kDa

Excitation / Emission

Unconjugated

Immunogen

Clone (M168) was generated from a mouse recombinant E-Cadherin protein containing amino acids in the cytoplasmic region. This sequence is highly conserved in human and rat E-cadherin.

Immunogen Species

Mouse

Species Reactivity

Human, Mouse, Rat

Storage

Aliquot and store at ≤ -20°C for long term storage. For short term storage, store at 2-8°C. For maximum recovery of product, centrifuge the vial prior to removing the cap.

Physical State

Liquid

Production Notes

This recombinant antibody is a chimeric antibody created by replacing the mouse heavy and light constant regions of clone M168 with chicken IgY heavy and light constant regions. As such this antibody retains the same binding performance as the original clone M168 but can be detected using standard anti-chicken secondary antibodies allowing flexibility for multiplexing applications. This antibody is expressed recombinantly in mammalian cells and then affinity purified from the cell culture media.

Buffer

1X PBS, 0.05% Sodium Azide pH 7.4

Dilution Ranges

WB: 1:1000-1:3000
IHC: 1:250-1:500
ICC: 1:250-1:1000
IP: 1:100

Conjugate

Unconjugated

Specificity

Cross-reacts with GFAP-R416W and other GFAP mutant proteins. Does not cross-react with other proteins (based on KO validation results).

Quality Control

Each new lot of antibody is quality control tested by Western blot on MCF-7 cell lysates and confirmed to stain the expected molecular weight band.

Usage Statement

These antibodies are to be used as research laboratory reagents and are not for use as diagnostic or therapeutic reagents in humans.

Country of Origin

United States

Expiration

24 months from date of receipt

Synonyms

AA960649; Uvomorulin; E-cad; L-CAM; E-cadherin; Cadherin-1; CTF1; CTF2; CTF3; CD324; Epithelial Cadherin

UniProt ID

P09803
UniProt Summary: Cadherins are calcium-dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells; cadherins may thus contribute to the sorting of heterogeneous cell types. CDH1 is involved in mechanisms regulating cell-cell adhesions, mobility and proliferation of epithelial cells. Promotes organization of radial actin fiber structure and cellular response to contractile forces, via its interaction with AMOTL2 which facilitates anchoring of radial actin fibers to CDH1 junction complexes at the cell membrane. Plays a role in the early stages of desmosome cell-cell junction formation via facilitating the recruitment of DSG2 and DSP to desmosome plaques. Has a potent invasive suppressor role. It is a ligand for integrin alpha-E/beta-7.

NCBI Gene ID

12550

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Anti-E-Cadherin (Cytoplasmic) Recombinant Chicken Chimeric mAb (M168)

SKU: 78-608-020

Product Details

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