Digoxigenin Antikörper

Produktnummer ABIN2473338

Zitiert in 1 Publikation. Der Schaf Polyklonal anti-Digoxigenin Antikörper (ABIN2473338) detektiert spezifisch Digoxigenin in ELISA, WB und IHC (fro). Dieser Antikörper reagiert spezifisch mit Proben aus Chemical. Er hat eine unabhängige Validierung für Immunohistochemistry.

Kurzübersicht für Digoxigenin Antikörper (ABIN2473338)

Target: Digoxigenin (DIG)

Reaktivität: Chemical

Wirt: Schaf

Klonalität: Polyklonal

Konjugat: Dieser Digoxigenin Antikörper ist unkonjugiert

Applikation: ELISA, Western Blotting (WB), Immunohistochemistry (Frozen Sections) (IHC (fro))

Produktdetails anti-Digoxigenin Antikörper

Produktmerkmale: Purified IgG

Aufreinigung: Purified

Immunogen: Digoxin-3-CM conjugated to keyhole limpet haemocyanin

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: Optimal working dilution should be determined by the investigator.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #104646 (Immunohistochemistry)

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

No.: #104646

Datum: 08.06.2026

Antigen: Digoxigenin (DIG)

Chargennummer: 171281

Validierte Anwendung: Immunohistochemistry

Positivkontrolle:

Sections from the same tissues were incubated with an anti-digoxigenin antibody conjugated with alkaline phosphatase (AP) and developed with NBT/BCIP solution.

Negativkontrolle:

Omission of Terminal transferase.

Bewertung:

The primary antibody works in IHC-P at concentrations from 1:200-1:500. The secondary antibody works in IHC-P at concentration of 1:1000.

Validierungsbilder

Positive control with Alkaline Phosphatase Anti-Digoxigenin antibody on P5 CD1 mouse brain. Images show reaction with (positive control) and without (TUNEL reaction) DNase treatment. Magnification: 40x.

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

Protokoll

Primärantikörper: ABIN2473338

Sekundärantikörper: ABIN102261 – Rabbit anti-Sheep IgG (Heavy & Light Chain) Antibody (FITC) – Preadsorbed (Rockland Immunochemicals, Inc.), lot# 27867

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 primary antibody at the following dilutions of 1:50, 1:200, 1:500, overnight at +4 °C (primary antibody was diluted in PLL/BSA/PBS diluent added with 5% BSA).
• 3 × 5-minute washes in 0.01 M PBS.
• Incubation with the anti-mouse secondary antibody 1:1000 in PLL/BSA/PBS diluent, 1 hour at RT.
• 3 × 5-minute washes 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.

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

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: 5 mg/mL

Referenzen für anti-Digoxigenin Antikörper (ABIN2473338)

Mori, Itoi, Tsukamoto, Kubo, Amagai: "The perivascular space as a path of hematopoietic progenitor cells and mature T cells between the blood circulation and the thymic parenchyma." in: International immunology, Vol. 19, Issue 6, pp. 745-53, (2007) (PubMed).

Details zu Digoxigenin

Target: Digoxigenin (DIG)

Andere Bezeichnung: DIGOXIGENIN

Substanzklasse: Chemical