PRODUCT CODE: EM0502

E-Cadherin Mouse Monoclonal Antibody [A0-G11-2] (EM0502)

  • IVD–IHC

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

All lanes: Western blot analysis of E-Cadherin with anti-E-Cadherin antibody [A0-G11-2] (EM0502) at 1:1,000 dilution.<br />
Lane 1/2: Wild-type A431 whole cell lysate (20 µg).<br />
Lane 3/4: E-Cadherin fragment 1 knockdown A431 whole cell lysate (20 µg).<br />
Lane 5/6: E-Cadherin fragment 2 knockdown A431 whole cell lysate (20 µg).<br />
<br />
EM0502 was shown to specifically react with E-Cadherin in wild-type A431 cells. Weakened bands were observed when E-Cadherin knockdown samples were tested. Wild-type and E-Cadherin knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (EM0502, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.
  • All lanes: Western blot analysis of E-Cadherin with anti-E-Cadherin antibody [A0-G11-2] (EM0502) at 1:1,000 dilution.<br />
Lane 1/2: Wild-type A431 whole cell lysate (20 µg).<br />
Lane 3/4: E-Cadherin fragment 1 knockdown A431 whole cell lysate (20 µg).<br />
Lane 5/6: E-Cadherin fragment 2 knockdown A431 whole cell lysate (20 µg).<br />
<br />
EM0502 was shown to specifically react with E-Cadherin in wild-type A431 cells. Weakened bands were observed when E-Cadherin knockdown samples were tested. Wild-type and E-Cadherin knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (EM0502, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of E-Cadherin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM0502, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: A431 cell lysate<br />
Lane 2: SW480 cell lysate
  • ICC staining of E-Cadherin in A431 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (EM0502, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®555 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
  • Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-E-Cadherin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0502, 1/50) for 30 minutes 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.
  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-E-Cadherin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM0502, 1/50) for 30 minutes 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.
  • Flow cytometric analysis of E-Cadherin was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (EM0502, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
All lanes: Western blot analysis of E-Cadherin with anti-E-Cadherin antibody [A0-G11-2] (EM0502) at 1:1,000 dilution.
Lane 1/2: Wild-type A431 whole cell lysate (20 µg).
Lane 3/4: E-Cadherin fragment 1 knockdown A431 whole cell lysate (20 µg).
Lane 5/6: E-Cadherin fragment 2 knockdown A431 whole cell lysate (20 µg).

EM0502 was shown to specifically react with E-Cadherin in wild-type A431 cells. Weakened bands were observed when E-Cadherin knockdown samples were tested. Wild-type and E-Cadherin knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (EM0502, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

E-Cadherin Mouse Monoclonal Antibody [A0-G11-2] (EM0502)

Immunogen

Recombinant protein

Host

Mouse

Positive Control

A431 cell lysate, SW480 cell lysate, A431, human liver carcinoma tissue, human colon carcinoma tissue, Hela.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

A0-G11-2

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

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

Concentration

2 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

130 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:500-1:1,000

  • ICC

  • 1:200

  • IHC-P

  • 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; ECAD antibody; Epithelial cadherin antibody; epithelial calcium dependant adhesion protein antibody; LCAM antibody; Liver cell adhesion molecule antibody; UVO antibody; Uvomorulin antibody

TISSUE SPECIFICITY

Expressed in inner and outer pillar cells of the organ of Corti (at protein level). Non-neural epithelial tissues.

DEVELOPMENTAL STAGE

In the testis, expression is highest in fetal gonad, then decreases 5-fold in newborn. Detectable in 7-day-old but not in 21-day-old or adult.

POST-TRANSLATIONAL MODIFICATION

During apoptosis or with calcium influx, cleaved by a membrane-bound metalloproteinase (ADAM10), PS1/gamma-secretase and caspase-3 (By similarity). Processing by the metalloproteinase, induced by calcium influx, causes disruption of cell-cell adhesion and the subsequent release of beta-catenin into the cytoplasm (By similarity). The residual membrane-tethered cleavage product is rapidly degraded via an intracellular proteolytic pathway (By similarity). Cleavage by caspase-3 releases the cytoplasmic tail resulting in disintegration of the actin microfilament system (By similarity). The gamma-secretase-mediated cleavage promotes disassembly of adherens junctions (By similarity). During development of the cochlear organ of Corti, cleavage by ADAM10 at adherens junctions promotes pillar cell separation.; O-glycosylated. O-manosylated by TMTC1, TMTC2, TMTC3 or TMTC4. Ser-287 and Thr-511 are O-manosylated by TMTC2 or TMTC4 but not TMTC1 or TMTC3.; N-glycosylation at Asn-639 is essential for expression, folding and trafficking. Addition of bisecting N-acetylglucosamine by MGAT3 modulates its cell membrane location (By similarity).; Ubiquitinated by a SCF complex containing SKP2, which requires prior phosphorylation by CK1/CSNK1A1. Ubiquitinated by CBLL1/HAKAI, requires prior phosphorylation at Tyr-756 (By similarity).

SUBCELLULAR LOCATION

Cell membrane

FUNCTION

E-cadherin (epithelial) is the most well-studied member of the cadherin family. It consists of 5 cadherin repeats (EC1 ~ EC5) in the extracellular domain, one transmembrane domain, and an intracellular domain that binds p120-catenin and beta-catenin. The intracellular domain contains a highly-phosphorylated region vital to beta-catenin binding and, therefore, to E-cadherin function. Loss of E-cadherin function or expression has been implicated in cancer progression and metastasis. E-cadherin downregulation decreases the strength of cellular adhesion within a tissue, resulting in an increase in cellular motility. This in turn may allow cancer cells to cross the basement membrane and invade surrounding tissues. E-cadherin is also used by pathologists to diagnose different kinds of breast cancer.