TFF3 Antikörper (AA 23-81)

Produktnummer ABIN5692886

Produktdetails anti-TFF3 Antikörper

Target: TFF3 (Trefoil Factor 3 (Intestinal) (TFF3))

Bindungsspezifität: AA 23-81

Reaktivität: Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser TFF3 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), ELISA, Immunohistochemistry (IHC)

Marke: Picoband™

Kreuzreaktivität (Details): No cross reactivity with other proteins.

Produktmerkmale: Rabbit IgG polyclonal antibody for TFF3 detection. Tested with WB, IHC-P, Direct ELISA in Mouse,Rat.

Immunogen: E. coli-derived mouse TFF3 recombinant protein (Position: A23-F81).

Anwendungsinformationen

Applikationshinweise:

Recommended Detection Systems: Enhanced Chemiluminescent Kit with anti-Rabbit IgG (ABIN921124) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).

Application Details: Western blot, 0.1-0.5 μg/mL
Immunohistochemistry(Paraffin-embedded Section), 0.5-1 μg/mL
Direct ELISA, 0.1-0.5 μg/mL

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #103596 (Western Blotting)

by: Laboratory for neurodegenerative disease research, Department of Molecular Medicine, Ruđer Bošković Institute, Zagreb, Croatia

No.: #103596

Datum: 18.02.2019

Antigen: Tff3

Chargennummer: 0011712Da513858

Validierte Anwendung: Western Blotting

Positivkontrolle:

colon from WT C57BL/6 mice

Negativkontrolle:

small intestine from Tff3 deficient mice

Bewertung:

ABIN5692886 specifically recognizes Tff3 in colon and small intestine tissue of wild type C57BL/6 mice.

Validierungsbilder

ABIN5692886 specifically recognizes Tff3 in colon and small intestine tissue of wild type C57BL/6 mice.

Protokoll

Primärantikörper: ABIN5692886

Sekundärantikörper: goat anti-rabbit IgG-HRP antibody (Bio-Rad, 170-6515, L005679 A)

Full Protocol:

• Isolate colon and small intestine tissue of wild type C57BL/6 mice and small intestine of Tff3 deficient mice using RIPA buffer (50mM TRIS HCL, pH8, 150mM NaCl, 1mM EDTA, 1% NP40 (Sigma-Aldrich, I-3021), 1% sodium deoxycholate (Sigma-Aldrich, D6750-25g), 0.1% SDS) containing PhosSTOP phosphatase inhibitor (Roche, 04906845001, lot 27423300) and cOmplete Mini protease inhibitor (Roche, lot 25765800).
• Determine total protein concentration using Pierce BCA protein assay kit (ThermoFisher Scientific, 23227, lot Tf268093).
• Denature 30µg total protein for 5min at 95°C in 6µl 5x Laemmli SDS sample buffer, 3µl 3M DTT and 1.5ul β-mercaptoethanol and Milli-Q H₂0 in 30µl total volume per sample.
• Separate proteins on a freshly cast reducing 15% SDS-PAGE gel at 100V for the stacking gel and 150V for the running gel for 2-3 hours in a Mini-PROTEAN Tetra Vertical Electrophoresis Cell chamber (Bio-Rad).
• Transfer proteins onto a nitrocellulose membrane using Trans-Blot Transfer Medium (Bio-Rad, 162-0112, lot BR 8158009) on a Trans-Blot Turbo Transfer System (Bio-Rad, 1704150) at 150mA for 90min.
• Stain membrane using Ponceau S to verify protein transfer.
• Wash membrane with TBS.
• Block the membrane with 1x TBS, 5% non-fat Milk, 0.1% Tween) for 1h at RT.
• Incubate membrane with primary
• • polyclonal rabbit anti-TFF3 antibody (antibodies-online, ABIN5692886, lot 0011712Da513858) diluted 1:1000 in 1x TBS, 5% BSA, 0.1% Tween ON at 4°C.
  • polyclonal rabbit anti-TFF3 antibody (in-house) diluted in 1x TBS, 5% BSA, 0.1% Tween ON at 4°C.
• Wash membrane 3x for 10min with TBST (1x TBS, 0.1% Tween).
• Incubate membrane with secondary goat anti-rabbit IgG-HRP antibody (Bio-Rad, 170-6515, L005679 A) diluted 1: 5000 in TBST for 1.5h at RT.
• Wash membrane 3x for 10min with TBST.
• Reveal proteins bands using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, 32106, lot NJ175924) on an Imager Alliance 4.7 western blot imaging system (Uvitec).

Anmerkungen:

ABIN5692886 reveals a protein of the expected molecular weight of the antigen in mice tissue. The protein bands is only visible in the positive but not the negative control. The same protein band was also detected by an in-house anti-TFF3 antibody hat was used as a reference.

Handhabung

Format: Lyophilized

Rekonstitution: Add 0.2 mL of distilled water will yield a concentration of 500 μg/mL.

Buffer: Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na₂HPO₄, 0.05 mg NaN₃.

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: At -20°C for one year. After reconstitution, at 4°C for one month.
It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.

Details zu TFF3

Target: TFF3 (Trefoil Factor 3 (Intestinal) (TFF3))

Andere Bezeichnung: Tff3 (TFF3 Produkte)

Synonyme: TFF3 antikoerper, ITF antikoerper, P1B antikoerper, TFI antikoerper, mITF antikoerper, trefoil factor 3 antikoerper, trefoil factor 3, intestinal antikoerper, TFF3 antikoerper, Tff3 antikoerper

Hintergrund:

Synonyms: Trefoil factor 3, Intestinal trefoil factor, mITF, Tff3, Itf

Tissue Specificity: Expressed in goblet cells of the intestines and colon (at protein level). Expressed abundantly in goblet cells of intestine and colon, and at low levels in stomach. No expression in brain, lung, spleen, kidney, uterus, pancreas, liver, heart or thymus.

Background: Trefoil factor 3 is a protein that in humans is encoded by the TFF3 gene. It is mapped to 21q22.3. Members of the trefoil family can interact with mucins and have an influence on mucus viscosity. They also promote migration of epithelial cells, are linked to antiapoptosis, induce cell scattering, trigger chemotaxis, and participate in immune responses. This gene is a marker of columnar epithelium and is expressed in a variety of tissues including goblet cells of the intestines and colon. It has been found that TFF3 is the most highly upregulated gene in 19 poorly differentiated endometrioid endometrial carcinomas (G3-EECs) compared with normal endometrium biopsies.

UniProt: Q62395