PRODUCT CODE: 0407-25

E-Cadherin Antibody (0407-25)

Applications

  • WB

  • ICC

  • FC

REACTIVITY

  • Human

  • Mouse

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Western blot analysis of E-Cadherin on A431 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of E-Cadherin on A431 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining E-Cadherin in HCT116 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the antibody (0407-25) at a dilution of 1:200 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution.
  • ICC staining E-Cadherin in SKOV-3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the antibody (0407-25) at a dilution of 1:200 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution.
  • Flow cytometric analysis of E-Cadherin was done on Hela cells. The cells were fixed, permeabilized and stained with E-Cadherin antibody at 1/100 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.
Western blot analysis of E-Cadherin on A431 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • FC

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

E-Cadherin Antibody (0407-25)

Immunogen

This antibody is produced by immunizing rabbits with a synthetic peptide (klh-coupled) corresponding to a region of outside membrane of e-cadherin.

Host

Rabbit

Positive Control

HCT116, SKOV-3, Hela.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Peptide affinity purified.

MOLECULAR WEIGHT

80-124kDa

Isotype

IgG

APPLICATION DILUTION

  • WB:1:500

  • ICC:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

E-Cadherin

SYNONYMS

Arc 1 antibody; CADH1_HUMAN antibody; Cadherin 1 antibody; cadherin 1 type 1 E-cadherin antibody; Cadherin1 antibody; CAM 120/80 antibody; CD 324 antibody; CD324 antibody; CD324 antigen antibody; cdh1 antibody; CDHE antibody; E-Cad/CTF3 antibody; E Cadherin antibody; E-cadherin antibody; ECAD antibody; Epithelial cadherin antibody; epithelial calcium dependant adhesion protein antibody; LCAM antibody; Liver cell adhesion molecule antibody; UVO antibody; Uvomorulin antibody

TISSUE SPECIFICITY

Non-neural epithelial tissues.

POST-TRANSLATIONAL MODIFICATION

During apoptosis or with calcium influx, cleaved by a membrane-bound metalloproteinase (ADAM10), PS1/gamma-secretase and caspase-3. Processing by the metalloproteinase, induced by calcium influx, causes disruption of cell-cell adhesion and the subsequent release of beta-catenin into the cytoplasm. The residual membrane-tethered cleavage product is rapidly degraded via an intracellular proteolytic pathway. Cleavage by caspase-3 releases the cytoplasmic tail resulting in disintegration of the actin microfilament system. The gamma-secretase-mediated cleavage promotes disassembly of adherens junctions. During development of the cochlear organ of Corti, cleavage by ADAM10 at adherens junctions promotes pillar cell separation (By similarity).; N-glycosylation at Asn-637 is essential for expression, folding and trafficking. Addition of bisecting N-acetylglucosamine by MGAT3 modulates its cell membrane location.; Ubiquitinated by a SCF complex containing SKP2, which requires prior phosphorylation by CK1/CSNK1A1. Ubiquitinated by CBLL1/HAKAI, requires prior phosphorylation at Tyr-754.; O-glycosylated. O-manosylated by TMTC1, TMTC2, TMTC3 or TMTC4. Thr-285 and Thr-509 are O-mannosylated by TMTC2 or TMTC4 but not TMTC1 or TMTC3.

SUBCELLULAR LOCATION

Cell membrane.

FUNCTION

Cadherins are calcium-dependent cell adhesion proteins. The classic cadherin subfamily includes N-, P-, R-, B- and E-cadherins as well as about ten other members which are found in adherens junctions (AJ), E-cadherin is found at the junctions of epithelial cells. Besides mediating cell-cell adhesion through homophilic interactions, E-cadherin mediates contact inhibition of cell growth, and loss of E-cadherin is associated with tumorigenesis. E-cadherin expression is frequently down-regulated or extinguished in malignancy which strongly correlates with poor prognosis.

CITATIONS

  • Zhou, Yong-Chun et al.

    Ionizing radiation promotes migration and invasion of cancer cells through transforming growth factor-beta-mediated epithelial-mesenchymal transition. | International Journal of Radiation Oncology, Biology, Physics [2011]