Digoxigenin Antikörper (FITC)

Produktnummer ABIN1514934

Der FITC-konjugierte Maus Monoklonal anti-Digoxigenin Antikörper (Klon 21H8) (ABIN1514934) detektiert spezifisch Digoxigenin in IHC, ISH und FACS. Dieser Antikörper reagiert spezifisch mit Proben aus Chemical. Er hat eine unabhängige Validierung für Immunohistochemistry.

Kurzübersicht für Digoxigenin Antikörper (FITC) (ABIN1514934)

Target: Digoxigenin (DIG)

Reaktivität: Chemical

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser Digoxigenin Antikörper ist konjugiert mit FITC

Applikation: Immunohistochemistry (IHC), In situ hybridization (ISH), Flow Cytometry (FACS)

Klon: 21H8

Produktdetails anti-Digoxigenin Antikörper (FITC)

Spezifität: Reacts with free and bound digoxigenin

Aufreinigung: Purified (protein A)

Immunogen: Digoxigenin

Isotyp: IgG1 kappa

Anwendungsinformationen

Applikationshinweise:

Working dilution: Optimal dilution should be determined by the end user.
The following are guidelines only:
IHC-F(1:20 - 1:2000)

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #102325 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #102325

Datum: 08.06.2026

Antigen: Digoxigenin (DIG)

Chargennummer: 17F7

Validierte Anwendung: Immunohistochemistry

Positivkontrolle:

Alkaline Phosphatase Anti-Digoxigenin antibody (Abcam, Catalog # ab119345).

Negativkontrolle:

Omission of Terminal transferase.

Bewertung:

The antibody works in IHC-P at concentration of 1:100.

Validierungsbilder

Positive control with Alkaline Phosphatase Anti-Digoxigenin antibody on P5 CD1 mouse brain. Images show reaction with and without DNase treatment to increase cellular death. Magnification: 40x.

Digoxigenin (DIG) (FITC-conjugated) immunoreactive cells in the cerebral cortex, hippocampus and cerebellum of P5 CD1 mice. Sections were treated with DNase and then subjected to the TUNEL reaction. Magnification: 40x.

Protokoll

Primärantikörper: ABIN1514934

Full Protocol:

• Sample analyzed: P5 CD1 mouse brain
• Paraffin-embedded slices (7 µm thick) were deparaffinized and rehydrated through a graded series of alcohols and distilled water (dH₂O).
• Sections were incubated in proteinase K [20 µg/mL] for 15 minutes at room temperature (RT).
• 4 × 2 minutes washing in double distilled water (ddH₂O).
• Sections were briefly washed in DN buffer and incubated in DNase [1 µg/mL] for 10 minutes at RT.
• 4 × 2 minutes washing in ddH₂O.
• 5 minutes washing in TDT buffer at RT.
• Sections were incubated in TDT buffer added with Terminal transferase [0.05 U/µL] and digoxigenin-11-dUTP [10 µM] for 2 hours at +37 °C.
• 15 minutes washing in TB buffer at RT.
• 5 minutes washing in ddH₂O + 5 minutes washing in 0.01 M PBS.
• Sections were incubated with the FITC-conjugated antibody at the dilution of 1:100, overnight at +4 °C (antibody was diluted in PLL/BSA/PBS diluent added with 5% BSA).
• 3 × 5 minutes washing in 0.01 M PBS.
• Nuclear counterstaining was performed with 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI) [1 µg/mL], 2 minutes.
• 2 × 5 minutes washing in 0.01 M PBS.
• Specimens were mounted in Fluoroshield mounting medium (Sigma).
• Images were acquired with a Leica DM 6000B fluorescence microscope with a digital camera at 40× magnification.

• POSITIVE CONTROL: Sections from the same tissues were incubated with an anti-digoxigenin antibody conjugated with alkaline phosphatase (AP) and developed with NBT/BCIP solution.
• NEGATIVE CONTROL: Omission of Terminal transferase.
• OTHER PERTINENT REAGENTS USED:
  • Proteinase K (Thermo Scientific, Catalog # EO0491)
  • DNase I (in the kit by Sigma, Catalog # AMP-D1)
  • Terminal transferase (in the kit by Jena Biosciences, Catalog # APP-002)
  • Digoxigenin-11-dUTP (Roche, Catalog # 11573179910)
  • DN buffer: Tris base [30 mM] pH 7.2, sodium cacodylate [140 mM], magnesium chloride [4 mM], DTT [0.1 mM] in dH₂O
  • TDT buffer: Tris base [30 mM] pH 7.2, sodium cacodylate [140 mM], cobalt chloride [1 mM] in dH₂O
  • TB buffer: sodium chloride [300 mM], sodium citrate [30 mM] in dH₂O
  • For positive control: Alkaline Phosphatase Anti-Digoxigenin antibody (Abcam, Catalog # ab119345)

Anmerkungen:

To exclude any nonspecific signal due to the secondary antibody, one control slice was processed for each experimental procedure, omitting the primary antibody.

Handhabung

Format: Liquid

Konzentration: 0.2 mg/mL

Buffer: PBS, Glycerol 50 % , BSA 1 %,( pH 7.4), Thiomersal 0.02 %

Konservierungsmittel: Thimerosal (Merthiolate)

Vorsichtsmaßnahmen: This product contains Thimerosal (Merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: -20 °C

Informationen zur Lagerung: Store at -20°C. Avoid freeze-thaw cycles.

Details zu Digoxigenin

Target: Digoxigenin (DIG)

Andere Bezeichnung: Digoxigenin

Substanzklasse: Chemical

Hintergrund: Digoxigenin (DIG) is a hapten, a small molecule with high immunogenicity, that is used in many molecular biology applications.