Biotin antibody (DyLight 800)

Catalog No. ABIN6720668

Supplier Product No.: 600-445-098

The Rabbit Polyclonal anti-Biotin antibody is suitable to detect Biotin. It has been validated for WB, FM and FLISA. The antibody has been independently validated.

Quick Overview for Biotin antibody (DyLight 800) (ABIN6720668)

Target: Biotin

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Biotin antibody is conjugated to DyLight 800

Application: Western Blotting (WB), Fluorescence Microscopy (FM), FLISA

Product Details anti-Biotin Antibody (DyLight 800)

Supplier Product No.: 600-445-098

Supplier: Rockland

Purpose: Biotin Antibody Dylight™ 800 Conjugated

Cross-Reactivity (Details): Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum, Biotin conjugated IgG and Biotin conjugated Bovine Serum Albumin.

Purification: This product was prepared from monospecific antiserum by immunoaffinity chromatography using Biotin coupled to sepharose beads.

Immunogen: Biotin conjugated to Keyhole Limpet Hemocyanin (KLH)

Isotype: IgG

Labeling Ratio: 2.5

Application Details

Application Notes:

FLISA_Dilution: >1:20,000

IF_Microscopy_Dilution: >1:5,000

Western_Blot_Dilution: 1:10,000 - 1:25,000

Other: User Optimized

Comment:

Anti-Biotin DyLight 800 Conjugated Antibody has been tested by western blot. This product is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms. The emission spectra for this DyLight™ conjugate match the principle output wavelengths of most common fluorescence instrumentation.

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104645 (Immunohistochemistry)

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

No.: #104645

Date: 11/09/2026

Antigen: Biotin

Lot Number: 24716

Method validated: Immunohistochemistry

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.

Notes:

• The antibody works in IHC-P at a concentration of 1:500-1:5000.

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.

The TUNEL reaction was performed using biotinylated dUTP and revealed with ABIN6720668  (anti-biotin DyLight 800-conjugated). Magnification: 20x.

Full Methods

Primary Antibody: ABIN6720668

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

Experimental Notes:

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

Handling

Format: Lyophilized

Reconstitution:

Reconstitution_Buffer: Restore with deionized water (or equivalent)

Reconstitution_Volume: 100 μL

Buffer:

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Stabilizer: 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free

Preservative: 0.01 % (w/v) Sodium Azide

Preservative: Sodium azide

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

Storage: 4 °C,-20 °C

Storage Comment: Store vial at 4° C prior to restoration.   For extended storage aliquot contents and freeze at -20° C or below.  Avoid cycles of freezing and thawing.  Centrifuge product if not completely clear after standing at room temperature.  This product is stable for several weeks at 4° C as an undiluted liquid.  Dilute only prior to immediate use. 

Expiry Date: 12 months

Target Details for Biotin

Target: Biotin

Target Type: Chemical

Background:

Synonyms: rabbit anti-Biotin Antibody Dylight™ 800 Conjugated, rabbit anti Biotin Antibody Dylight™ 800 Conjugated

Background: Biotin Antibody detects Biotin. Biotin is a water-soluble B-complex vitamin (vitamin B7). It is composed of a ureido (tetrahydroimidizalone) ring fused with a tetrahydrothiophene ring. A valeric acid substituent is attached to one of the carbon atoms of the tetrahydrothiophene ring. Biotin is a coenzyme for carboxylase enzymes, involved in the synthesis of fatty acids, isoleucine, and valine, and in gluconeogenesis. Biotin is necessary for cell growth, the production of fatty acids, and the metabolism of fats and amino acids. Anti-Biotin DyLight Conjugated Antibody is ideal for investigators involved in Cell Signaling and Cell Biology research.