E-cadherin Antikörper (AA 375-631)

Produktnummer ABIN3209718

Produktdetails anti-E-cadherin Antikörper

Target: E-cadherin (CDH1) (Cadherin 1, Type 1, E-Cadherin (Epithelial) (CDH1))

Bindungsspezifität: AA 375-631

Reaktivität: Schwein

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser E-cadherin Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunocytochemistry (ICC)

Verwendungszweck: Polyclonal Antibody to E-cadherin

Spezifität: The antibody is a rabbit polyclonal antibody raised against E-cadherin. It has been selected for its ability to recognize E-cadherin in immunohistochemical staining and western blotting.

Aufreinigung: Antigen-specific affinity chromatography followed by Protein A affinity chromatography

Immunogen: Recombinant E-cadherin corresdonding to Phe375~Thr631 (Accession # C6EVT4) with N-terminal His and GST Tag

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise:

Western blotting: 0.5-2 μg/mL

1:160-660 Immunohistochemistry: 5-20 μg/mL

1:16-66 Immunocytochemistry: 5-20 μg/mL

1:16-66 Optimal working dilutions must be determined by end user.

Kommentare:

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IF)

Validierung #101345 (Immunofluorescence)

by: Sackler School of Biomedical Sciences, Tufts University School of Medicine

No.: #101345

Datum: 05.06.2017

Antigen: E-cadherin

Chargennummer: A20160427539

Validierte Anwendung: Immunofluorescence

Positivkontrolle: porcine unerupted tooth buds, dental epithelium

Negativkontrolle: porcine unerupted tooth buds, mesenchym

Bewertung: Passed. The E-caherin antibody ABIN1858334 specifically labels epithelial tissues in porcine unerupted tooth buds in immunofluorescence, consistent with the expression pattern of E-cadherin.

Validierungsbilder

A. ABIN1858334 stains the dental epithelium (DE, red) in pig tooth buds whereas the dental follicle (DF) and the pulp – labeled with the mesenchymal marker Vimentin (green) – remain unstained. B. Staining of the dental epithelial root tissue with ABIN1858334. C. Immunostaining of the DE with ABIN1858334. No staining of of dental mesenchym (DM) by ABIN1858334 or of the dental epithelium when using the secondary antibody alone (panle D). 40X, Scale Bar 50μm, 150 Resolution

Protokoll

Primärantikörper: ABIN1858334

Sekundärantikörper: goat anti-rabbit AF568 conjugated antibody (ThermoFisher Scientific, A11011, lot 1778025)

Full Protocol:

• Harvest unerupted tooth buds from 5 month old pig jaws and wash 3x in phosphate bufferend saline (PBS) (Boston Bioproducts, BM-220).
• Fix tooth buds in 10% Formalin (Fisher Scientific, 23-245-685) for 24h.
• Wash 3x in PBS.
• Place tooth buds through graded ethanol seris for 6h at each concentration: 50%, 70%, 80%, 90%, 100%, 100%, 100%.
• Place tooth buds in 3x 2h in 100% xylene.
• Place in molten paraffin 2x (TissuePrep 2 Paraffin, Fisher, T555-2) 12h.
• Embed tooth buds in paraffin and cool on a 4°C cold plate for 1h.
• Let set ON at RT.
• Cut blocks into 6µm sections using a microtome (ThermoFisher Scientific, HM 355S).
• Use a tissue floating bath to mount sections onto microscope slides.
• Heat sections on a hot plate for 1h at 45°C for 1h then ON at 55°C.
• Deparaffinize slide 2x 5 min in 100% xylene.
• Rehydrate sections in graded ethanol series for 2min for each concentration: 100%, 95%, 70%, 50%.
• Place slide in H₂O for 5min.
• Transfer slide to a plastic holder filled with warmed 10mM Citric Acid Solution (made from sodium citrate monobasic; Sigma, 71497) pH6.0 and incubate in a steamer for 20min.
• Cool submerged slide on benchtop for 30min.
• Wash slide 3x 5min in PBS.
• Create a hydrophobic barrier on slide around sections with PAP-pen (Vector Labs, H-4000, lot 5138).
• Place slide in a humidified chamber and block with 5% BSA for 20min at 37°C.
• Gently remove BSA by tapping slide.
• Incubate sections with primary antibody
• • rabbit anti-pig E-cadherin antibody (antibodies-online, ABIN1858334, lot A20160408626) diluted to 1:40 in 1% BSA or
  • and mouse anti-pig Vimentin (Santa Cruz Biotechnology, SC-6260, lot G2814) diluted to 1:500 in 1% BSA for 1h at RT.
• A no primary antibody negative control was incubated in parallel in 1% BSA.
• Wash in slide 3x 5min in PBS.
• Incubate sections in secondary antibodies
• • goat anti-rabbit AF568 conjugated antibody (ThermoFisher Scientific, A11011, lot 1778025) diluted to 1:100 in 1% BSA and
  • goat anti-mouse AF488 conjugated antibody (ThermoFisher Scientific, A11001, lot 1752514 ) diluted to 1:100 in 1% BSA for 1h at RT.
• Wash in slide 3x 5min in PBS.
• Add approximately 10μL of Hard Set Mounting Medium with DAPI (Vector Labs, H-1500, lot ZC1221) for each section and mount cover slip.

Handhabung

Format: Liquid

Konzentration: Lot specific

Buffer: PBS, pH 7.4, containing 0.02 % Sodium azide, 50 % glycerol.

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Store at 4°C for frequent use. Stored at -20°C in a manual defrost freezer for two year without detectable loss of activity. Avoid repeated freeze-thaw cycles.

Haltbarkeit: 24 months

Referenzen für anti-E-cadherin Antikörper (ABIN3209718)

Smith, Zhang, Schiele, Khademhosseini, Kuo, Yelick: "Developing a biomimetic tooth bud model." in: Journal of tissue engineering and regenerative medicine, (2017) (PubMed).

Monteiro, Smith, Angstadt, Zhang, Khademhosseini, Yelick: "Dental cell sheet biomimetic tooth bud model." in: Biomaterials, Vol. 106, pp. 167-79, (2016) (PubMed).

Details zu E-cadherin

Target: E-cadherin (CDH1) (Cadherin 1, Type 1, E-Cadherin (Epithelial) (CDH1))

Andere Bezeichnung: E-cadherin (CDH1 Produkte)

Synonyme: Arc-1 antikoerper, CD324 antikoerper, CDHE antikoerper, ECAD antikoerper, LCAM antikoerper, UVO antikoerper, E-cadherin antikoerper, cdh1 antikoerper, si:dz180o5.2 antikoerper, uvo antikoerper, lcam antikoerper, E-Cad antikoerper, XBcad antikoerper, l-cam antikoerper, xcdh1 antikoerper, cdhc-A antikoerper, xb-cad antikoerper, XTCAD-1 antikoerper, c-cadherin antikoerper, cadherin-1 antikoerper, uvomorulin antikoerper, XB-cadherin antikoerper, arc-1 antikoerper, cd324 antikoerper, cdhe antikoerper, ecad antikoerper, AA960649 antikoerper, E-cad antikoerper, Ecad antikoerper, L-CAM antikoerper, Um antikoerper, cadherin 1 antikoerper, si:busm1-180o5.2 antikoerper, cadherin 1, type 1, E-cadherin (epithelial) antikoerper, cadherin 1, type 1 S homeolog antikoerper, CDH1 antikoerper, si:busm1-180o5.2 antikoerper, cdh1 antikoerper, Cdh1 antikoerper, cdh1.S antikoerper

Hintergrund: CDHE, CD324, CDH1, Arc1, ECAD, CDH-E, E-CAD, LCAM, UVO, Cadherin 1 Type 1, Uvomorulin, E-Cadherin, Calcium-Dependent Adhesion Protein, Epithelial, Liver Cell Adhesion Molecule

Pathways: WNT Signalweg, Sensory Perception of Sound, Cell-Cell Junction Organization, Tube Formation