Digoxigenin anticorps (FITC)

N° du produit ABIN1514934

Cet anticorps anti-Digoxigenin Monoclonal Souris conjugué à FITC (Clone 21H8) (ABIN1514934) détecte spécifiquement Digoxigenin dans IHC, ISH et FACS. L’anticorps est réactif avec des échantillons de Chemical. Validé indépendamment pour une utilisation en Immunohistochemistry.

Aperçu rapide pour Digoxigenin anticorps (FITC) (ABIN1514934)

Antigène: Digoxigenin (DIG)

Reactivité: Chemical

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp Digoxigenin est conjugé à/à la FITC

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

Clone: 21H8

Détail du produit anti-Digoxigenin anticorps (FITC)

Specificité: Reacts with free and bound digoxigenin

Purification: Purified (protein A)

Immunogène: Digoxigenin

Isotype: IgG1 kappa

Information d'application

Indications d'application:

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

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #102325 (Immunohistochemistry)

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

No.: #102325

Date: 08.06.2026

Antigène: Digoxigenin (DIG)

Numéro du lot: 17F7

Application validée: Immunohistochemistry

Contrôle positif:

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

Contrôle négative:

Omission of Terminal transferase.

Conclusion:

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

Validation Images

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.

Protocole

Anticorps primaire: 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)

Notes:

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

Stockage

Format: Liquid

Concentration: 0.2 mg/mL

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

Agent conservateur: Thimerosal (Merthiolate)

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

Stock: -20 °C

Stockage commentaire: Store at -20°C. Avoid freeze-thaw cycles.

Détails sur Digoxigenin

Antigène: Digoxigenin (DIG)

Autre désignation: Digoxigenin

Classe de substances: Chemical

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