製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to EpCAM
  • アプリケーション
    適用あり: ICC/IF, IHC-P, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Dog, Human
    交差が予測される動物種: Cow, Pig, Rhesus monkey
  • 免疫原

    Synthetic peptide within Human EpCAM aa 250 to the C-terminus (internal sequence) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab71915)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following human lysates: HCT 116 Whole Cell, HepG2 Whole Cell, SW480 Whole Cell and Human Ovary Tissue. IHC-P: FFPE human breast adenocarcinoma tissue sections.

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab71916 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 1 - 5 µg/ml.
IHC-P 1/40 - 1/160.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).

ターゲット情報

  • 機能
    May act as a physical homophilic interaction molecule between intestinal epithelial cells (IECs) and intraepithelial lymphocytes (IELs) at the mucosal epithelium for providing immunological barrier as a first line of defense against mucosal infection. Plays a role in embryonic stem cells proliferation and differentiation. Up-regulates the expression of FABP5, MYC and cyclins A and E.
  • 組織特異性
    Highly and selectively expressed by undifferentiated rather than differentiated embryonic stem cells (ESC). Levels rapidly diminish as soon as ESC's differentiate (at protein levels). Expressed in almost all epithelial cell membranes but not on mesodermal or neural cell membranes. Found on the surface of adenocarcinoma.
  • 関連疾患
    Defects in EPCAM are the cause of diarrhea type 5 (DIAR5) [MIM:613217]. It is an intractable diarrhea of infancy characterized by villous atrophy and absence of inflammation, with intestinal epithelial cell dysplasia manifesting as focal epithelial tufts in the duodenum and jejunum.
    Defects in EPCAM are a cause of hereditary non-polyposis colorectal cancer type 8 (HNPCC8) [MIM:613244]. HNPCC is a disease associated with marked increase in cancer susceptibility. It is characterized by a familial predisposition to early-onset colorectal carcinoma (CRC) and extra-colonic tumors of the gastrointestinal, urological and female reproductive tracts. HNPCC is reported to be the most common form of inherited colorectal cancer in the Western world. Clinically, HNPCC is often divided into two subgroups. Type I is characterized by hereditary predisposition to colorectal cancer, a young age of onset, and carcinoma observed in the proximal colon. Type II is characterized by increased risk for cancers in certain tissues such as the uterus, ovary, breast, stomach, small intestine, skin, and larynx in addition to the colon. Diagnosis of classical HNPCC is based on the Amsterdam criteria: 3 or more relatives affected by colorectal cancer, one a first degree relative of the other two; 2 or more generation affected; 1 or more colorectal cancers presenting before 50 years of age; exclusion of hereditary polyposis syndromes. The term 'suspected HNPCC' or 'incomplete HNPCC' can be used to describe families who do not or only partially fulfill the Amsterdam criteria, but in whom a genetic basis for colon cancer is strongly suspected. Note=HNPCC8 results from heterozygous deletion of 3-prime exons of EPCAM and intergenic regions directly upstream of MSH2, resulting in transcriptional read-through and epigenetic silencing of MSH2 in tissues expressing EPCAM.
  • 配列類似性
    Belongs to the EPCAM family.
    Contains 1 thyroglobulin type-1 domain.
  • 翻訳後修飾
    Hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. Glycosylation at Asn-198 is crucial for protein stability.
  • 細胞内局在
    Lateral cell membrane. Cell junction > tight junction. Co-localizes with CLDN7 at the lateral cell membrane and tight junction.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 17 1A antibody
    • 323/A3 antibody
    • Adenocarcinoma associated antigen antibody
    • Adenocarcinoma-associated antigen antibody
    • Antigen identified by monoclonal antibody AUA1 antibody
    • AUA1 antibody
    • CD326 antibody
    • CD326 antigen antibody
    • Cell surface glycoprotein Trop 1 antibody
    • Cell surface glycoprotein Trop 2 antibody
    • Cell surface glycoprotein Trop-1 antibody
    • CO 17A antibody
    • CO17 1A antibody
    • CO17A antibody
    • DIAR5 antibody
    • EGP 2 antibody
    • EGP antibody
    • EGP2 antibody
    • EGP314 antibody
    • EGP40 antibody
    • Ep CAM antibody
    • Ep-CAM antibody
    • EPCAM antibody
    • EPCAM_HUMAN antibody
    • EpCAM1 antibody
    • Epithelial cell adhesion molecule antibody
    • Epithelial Cell Adhesion Molecule Intracellular Domain (EpCAM-ICD) antibody
    • Epithelial cell surface antigen antibody
    • Epithelial cellular adhesion molecule antibody
    • Epithelial glycoprotein 1 antibody
    • Epithelial glycoprotein 314 antibody
    • Epithelial glycoprotein antibody
    • ESA antibody
    • GA733 1 antibody
    • GA733 2 antibody
    • GA733-2 antibody
    • gastrointestinal tumor-associated antigen 2, 35-KD glycoprotein antibody
    • gp4 antibody
    • hEGP 2 antibody
    • hEGP314 antibody
    • HNPCC8 antibody
    • Human epithelial glycoprotein 2 antibody
    • KS 1/4 antigen antibody
    • KS1/4 antibody
    • KSA antibody
    • Ly74 antibody
    • Lymphocyte antigen 74 antibody
    • M1S 1 antibody
    • M1S2 antibody
    • M4S1 antibody
    • Major gastrointestinal tumor associated protein GA733 2 antibody
    • Major gastrointestinal tumor-associated protein GA733-2 antibody
    • mEGP314 antibody
    • Membrane component chromosome 4 surface marker (35kD glycoprotein) antibody
    • Membrane component, chromosome 4, surface marker 1 antibody
    • Membrane component, chromosome 4, surface marker antibody
    • MIC18 antibody
    • MK 1 antibody
    • Protein 289A antibody
    • TACD1 antibody
    • TACSTD1 antibody
    • TROP1 antibody
    • Tumor associated calcium signal transducer 1 antibody
    • Tumor associated calcium signal transducer 2 precursor antibody
    • Tumor-associated calcium signal transducer 1 antibody
    see all

画像

  • IHC image of EpCAM staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75962, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ICC/IF image of ab71916 stained Hek293 cells. The cells were 100% Methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab71916, 1µg/ml) overnight at +4°C. The secondary antibody (green) was goat anti rabbit Alexa Fluor® 488 (IgG H+L;  ab150077) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% Methanol fixed (5 min) HepG2, and MCF-7 cells at 5µg/ml, and in 4% PFA fixed (10 min) Hek293, HepG2, MCF-7 cells at 5µg/ml.

  • All lanes : Anti-EpCAM antibody (ab71916) at 1 µg/ml

    Lane 1 : Colon (Mouse) Tissue Lysate
    Lane 2 : Colon (Rat) Tissue Lysate
    Lane 3 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate
    Lane 4 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate
    Lane 5 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
    Lane 6 : HuES7 (Human embryonic stem cell line) Whole Cell Lysate
    Lane 7 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 35 kDa
    Observed band size : 35 kDa
    Additional bands at : 40 kDa. We are unsure as to the identity of these extra bands.

    Secondary antibody - goat anti-rabbit HRP (H&L preadsorbed;  ab97080)

  • ab71916 (1:160) staining EpCAM in paraffin-embedded human breast tissue, using an automated system (Ventana Discovery).
    Using this protocol there is strong membrane staining of the basolateral membranes of normal breast epithelial cells of breast ducts and lobular acini. There is associated weak to moderate staining of the cytoplasm of these cells.

    Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Standard Retrieval programme. Slides were blocked in 3% H2O2 / 4 min / 37°C and incubated with ab71916 (1:160 dilution / 2 hours / 37°C). Sections then blocked (4mins / 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min / 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min / 37°C) and Ventana DAB map reagent (8 min / 37°C). Slides were counterstained with Haematoxylin and coverslipped in DPX.

    For manua

  • Immunocytochemical immunofluorescence analysis of methanol-fixed rat primary alveolar type II cells, labelling EpCAM with ab71916 at at a dilution of 1/100 incubated for 1 hour at 25°C. Permeabilization was with Triton X. Blocking was with 5% BSA incubated for 30 minutes at 25°C. Secondary was a donkey anti-rabbit Alexa Fluor® 568 conjugate at 1/400.

    See Abreview

  • ab71916 (1:40) staining EpCAM in paraffin-embedded human breast carcinoma (Grade 2 Invasive Ductal Carcinoma) using an automated system (Ventana Discovery).
    Using this protocol there is moderate to strong membrane staining in carcinoma cells which may be apical or complete instead of basolateral. There is associated moderate to strong staining of the cytoplasm of these cells.

    Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Standard Retrieval programme. Slides were blocked in 3% H2O2 / 4 min / 37°C and incubated with ab71916 (1:40 dilution / 1 hour / 37°C). Sections then blocked (4mins / 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min / 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min / 37°C) and Ventana DAB map reagent (8 min / 37°C). Slides were counterstained with Haematoxylin and coverslipped in D

参考文献

This product has been referenced in:
  • Rengasamy M  et al. Transplantation of human bone marrow mesenchymal stromal cells reduces liver fibrosis more effectively than Wharton's jelly mesenchymal stromal cells. Stem Cell Res Ther 8:143 (2017). IHC-P ; Rat . Read more (PubMed: 28610623) »
  • Shih SC  et al. Pigment epithelium-derived factor (PEDF) peptide promotes the expansion of hepatic stem/progenitor cells via ERK and STAT3-dependent signaling. Am J Transl Res 9:1114-1126 (2017). Read more (PubMed: 28386338) »

See all 20 Publications for this product

レビューと Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Duodenum)
Specification
Duodenum
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Jack Robertson

Verified customer

投稿 Sep 18 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Ear wound exudates)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
No
Specification
Ear wound exudates
Blocking step
Protein Block serum free from Dako as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 37°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 12 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Mouse Cell (E9.5 embryo)
Permeabilization
No
Gating Strategy
same population, secondary only
Specification
E9.5 embryo
Preparation
Cell harvesting/tissue preparation method: Dissect and dissociate with trypsin
Sample buffer: HBSS +2% NGS +2% BSA
Fixation
no fixative
Username

Abcam user community

Verified customer

投稿 May 30 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 11 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (undifferentiated Human ES cells on MEFs)
Permeabilization
Yes - see blocking buffer
Specification
undifferentiated Human ES cells on MEFs
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Feb 22 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HUES7 ES cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
HUES7 ES cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Username

Abcam user community

Verified customer

投稿 Feb 04 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEPG2)
Specification
HEPG2
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Nov 18 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (HEPG2)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
HEPG2
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Username

Abcam user community

Verified customer

投稿 Nov 13 2015

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Hes3 Stem cell)
Permeabilization
No
Specification
Hes3 Stem cell
Username

Abcam user community

Verified customer

投稿 Oct 21 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary alveolar type II cells)
Permeabilization
Yes - Triton-X
Specification
Primary alveolar type II cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Methanol
Username

Abcam user community

Verified customer

投稿 Oct 19 2015

1-10 of 17 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

登録