E-Cadherin Antibody (SY0287)

Novus Biologicals | Catalog # NBP3-32290

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # SY0287 expressed in HEK293
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Product Specifications

Immunogen

Synthetic peptide within human E-Cadherin aa 580-630 (Extracellular). (Uniprot: P12830)

Localization

Cell junction, Cell membrane, Endosome, Golgi apparatus.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

97 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for E-Cadherin Antibody (SY0287)

E-Cadherin Antibody (SY0287)

Immunohistochemistry: E-Cadherin Antibody (SY0287) [NBP3-32290]

Immunohistochemistry: E-Cadherin Antibody (SY0287) [NBP3-32290] - Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Rabbit anti-E-Cadherin antibody (NBP3-32290) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32290) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
E-Cadherin Antibody (SY0287)

Immunohistochemistry: E-Cadherin Antibody (SY0287) [NBP3-32290]

Immunohistochemistry: E-Cadherin Antibody (SY0287) [NBP3-32290] - Immunofluorescence analysis of paraffin-embedded human colon tissue labeling E-Cadherin with Rabbit anti-E-Cadherin antibody (NBP3-32290) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (NBP3-32290, green) at 1/50 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with 33258 (blue).
E-Cadherin Antibody (SY0287)

Immunocytochemistry/ Immunofluorescence: E-Cadherin Antibody (SY0287) [NBP3-32290]

Immunocytochemistry/ Immunofluorescence: E-Cadherin Antibody (SY0287) [NBP3-32290] - Immunocytochemistry analysis of HT-29 (positive) and MDA-MB-231 (negative) cells labeling E-Cadherin with Rabbit anti-E-Cadherin antibody (NBP3-32290) at 1/2,000 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-E-Cadherin antibody (NBP3-32290) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
E-Cadherin Antibody (SY0287)

Flow Cytometry: E-Cadherin Antibody (SY0287) [NBP3-32290]

Flow Cytometry: E-Cadherin Antibody (SY0287) [NBP3-32290] - Flow cytometric analysis of HT-29 (positive, red) and MDA-MB-231 (negative, green) cells labeling E-Cadherin.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32290, red) at 1/1,000 dilution, compared with Rabbit IgG Isotype Control (HT-29 black, MDA-MB-231 light green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with an iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃.
E-Cadherin Antibody (SY0287)

Western Blot: E-Cadherin Antibody (SY0287) [NBP3-32290]

Western Blot: E-Cadherin Antibody (SY0287) [NBP3-32290] - Western blot analysis of E-Cadherin on different lysates with Rabbit anti-E-Cadherin antibody (NBP3-32290) at 1/5,000 dilution.
Lane 1: MCF7 cell lysate
Lane 2: MDA-MB-231 cell lysate (negative)
Lane 3: HT-29 cell lysate
Lane 4: HCT 116 cell lysate
Lane 5: A431 cell lysate
Lane 6: Caco-2 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 97 kDa
Observed band size: 80~120 kDa

Exposure time: 43 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.

Applications for E-Cadherin Antibody (SY0287)

Application
Recommended Usage

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

1:2000-1:200

Immunohistochemistry-Paraffin

1:200

Immunoprecipitation

Use at an assay dependent concentration

Western Blot

1:5000

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Advanced Features

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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

1*TBS (pH7.4), 0.05% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: E-Cadherin

E-cadherin, or epithelial cadherin, is an cell-cell adhesion glycoprotein important to cellular processes such as morphology, polarity, development, tissue integrity and migration. E cadherin's extracellular domain interacts with other E-cadherin protein on adjacent epithelial cells to establish adhesion between them. E cadherin has also been identified as a tumor and metastasis suppressor, and its mutations have been linked to various forms of cancer.

Alternate Names

Arc-1, CAD1, Cadherin-1, CD324, CDH1, Cell-CAM120/80, ECAD, ECadherin, L-CAM, Uvomorulin

Gene Symbol

CDH1

Additional E-Cadherin Products

Product Documents for E-Cadherin Antibody (SY0287)

Certificate of Analysis

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Product Specific Notices for E-Cadherin Antibody (SY0287)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

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Associated Pathways

Dendritic Cell Lineage Development Pathways Dendritic Cell Lineage Development Pathway Thumbnail