AccuSignal™ Nuclease ELISA Kit for Impurity Detection

N° du produit ABIN7505815

Détail du produit

Highlights:

• Détection d'impuretés dans la fabrication de médicaments biologiques.
• Le kit AccuSignal™ Nuclease ELISA est conçu pour une quantification sensible et fiable des nucléases dans les produits thérapeutiques, y compris DENARASE®, Benzonase® et Turbonuclease.
• Il offre un large spectre de quantification, tout en conservant une linéarité de dilution exceptionnelle, ce qui permet d'avoir confiance dans la précision des résultats sur un large éventail de concentrations de nucléases.
• Les résultats sont cohérents et reproductibles, caractérisés par une variabilité minimale intra- et inter-essais.
• La spécificité de l'anticorps permet une application sur divers matériaux, s'adaptant à divers produits de nucléases provenant de plusieurs fournisseurs.
• Composants : Plaque à 96 puits, scelleur de plaque, nucléase standard, anticorps de détection biotinylé, Streptavidine-HRO (100X), tampons.

Antigène: Nuclease

Méthode de détection: Colorimetric

Hôte: Lapin

Clonalité: Polyclonal

Type de méthode: Sandwich ELISA

Gamme de detection: 0.03 ng/mL - 20 ng/mL

Seuil minimal de détection: 0.03 ng/mL

Application: ELISA, Impurity Detection (Imp De)

Fonction: Nuclease ELISA kit is designed for the quantitative detection of Nuclease/NucA in serum, plasma, and hybridoma cell supernatants.

Marque: AccuSignal™

Analytical Method: Quantitative

Specificité: Benzonase, Nuclease, Endonuclease, Serratia marcescens endonuclease, NucA, Denarase

Sensibilité: 3 ng/mL

Ingrédients: This kit contains: Nuclease antibody coated 96-well strip plate, Plate Sealer, Nuclease standard, biotinylated Detection Antibody, streptavidin-peroxidase conjugate, along with buffers and protocol.

Matériel non inclus:

• Microplate shaker (up tp 450 rpm)
• Interval timer
• Multichannel pipettor (50-300 µL)
• Precision single pipettes (10 µL, 35 µL, 100 µL, 1000 µL, etc.)
• Disposable pipette tips
• Deionized water
• Disposable microcentrifuge Tube(s) or microplate
• Polypropylene centrifuge tubes (15 mL)
• Spectrophotometer microplate reader (450 nm absorbance, 630–650 nm reference filter)
• Disposable gloves
• Graduated cylinder
• Reagent reservoirs
• Vortex mixer
• Stir plate & magnetic stir bar
• Absorbent paper

Informations sur le produit:

Le kit ELISA pour Nucléases est un outil essentiel spécialement conçu pour quantifier de manière sensible et robuste la quantité de nucléases dans les produits thérapeutiques, y compris DENARASE®, Benzonase® et Turbonuclease. Ce kit utilise un format ELISA sandwich, exploitant la spécificité et l'affinité des anticorps anti-nucléase pré-immobilisés. La détection conjuguée à la biotine combinée avec la streptavidine-HRP garantit une sensibilité inégalée, permettant une mesure précise des nucléases dans une variété de matériaux thérapeutiques. Il facilite l'évaluation de la teneur en nucléases dans les thérapeutiques dérivées biologiquement, assurant leur stabilité et qualité tout au long du processus de fabrication. De plus, il aide à évaluer les nucléases dans les vecteurs de thérapie génique, un facteur critique pour maintenir l'intégrité de ces vecteurs pour une livraison génique thérapeutique efficace. Il aide également à valider les niveaux de nucléases dans les formulations pharmaceutiques, protégeant leur puissance et prévenant la dégradation potentielle. En fournissant une méthode fiable et standardisée pour la quantification des nucléases, le kit ELISA pour Nucléases assure non seulement la conformité avec des exigences réglementaires strictes mais accélère également les processus de recherche et développement.

Principaux domaines d'application du kit ELISA AccuSignal™ pour Nucléases

Thérapeutiques Biologiques : Évaluer les niveaux de nucléases dans les thérapeutiques biologiques pour garantir leur stabilité et qualité pendant la production.

Vecteurs de Thérapie Génique : Aider à évaluer la présence de nucléases dans les vecteurs de thérapie génique, essentiels pour préserver leur intégrité pour des traitements de livraison génique réussis.

Production de Vaccins : Confirmer les concentrations de nucléases dans les formulations pharmaceutiques pour assurer leur efficacité et prévenir la dégradation potentielle.

L'approche conviviale et la méthodologie robuste simplifient les flux de travail, permettant aux scientifiques de se concentrer sur l'avancement des innovations thérapeutiques en toute confiance. En tant qu'instrument indispensable, il favorise la précision, la fiabilité et l'efficacité dans l'évaluation des nucléases dans les produits thérapeutiques, tout en assurant la sécurité et l'efficacité.

Information d'application

Indications d'application: This kit was shown to positively detect commercially available nuclease variants, such as Benzonase, Denarase and other analogs.

Durée du test: 3 h

Préparation des réactifs:

Preparation of Standards & Test Samples

1. Reconstitute the standard vial with 1.0 mL deionized water to obtain a final concentration of 200 ng/mL.
Note: This is the “reference stock solution” that will be used below to make the standards.
2. Prepare dilutions of standard.
3. Test samples should be diluted in Nuclease Kit Sample Buffer based on empirically determined criteria for each sample.

Detection antibody working solution preparation

1. Add 110 µL of conjugated antibody to 11 mL of Sample Buffer for use in a full 96-well assay.
2. Mix well by pipette or inversion. Do not vortex.
3. Distribute antibody working solution as described in the assay procedure.

Note: Volumes may be adjusted so long as final working concentration remains as specified.

Streptavidin-HRP Working Solution Preparation

1. Add 110 µL of Streptavidin-HRP to 11 mL of Sample Buffer, respectively for use in a full 96-well assay.
2. Mix well by pipette or inversion. Do not vortex.
3. Distribute Streptavidin-HRP working solution as described in the assay procedure.

Note: Volumes may be adjusted so long as final working concentration remains as specified

Wash Buffer (1X) Preparation

1. Add 50 mL of Nuclease Kit Wash Buffer (10X) to 450 mL of deionized water.
2. Mix for at least 10-minutes using a magnetic stir bar.

Note: The wash buffer (1X) can be stored at room temperature (15°C to 25°C) for up to 2-weeks, after which it should be discarded.

Procédure de l'essai:

1. To each well add 100 µL of unknown or standard sample per well and incubate at room temperature for 60-minutes with shaking at 450 revolutions per minutes (rpm) on a shaker.
2. Wash the wells with Wash Buffer as follows:
• Decant the contents of the wells manually with a hard, rapid downward motion. Fluid should be captured in a receptable designed to collect the waste. Remove all residual reagent from the microplate by tapping it on absorbent paper with the opening facing downwards.
• Fill each well with 300 µL of Washing Buffer with a multichannel pipettor.
• Decant the Washing Buffer from the wells with a hard, rapid downward motion. Remove all residual solution from the microplate by tapping it on absorbent paper with the opening facing downwards.
• Repeat steps ii and iii two more times (total of 3 washings). Do not leave any residual moisture in the wells on each washing step.
3. To each well add 100 µL of detection antibody working solution
4. Incubate at room temperature for 60-minutes, covered to protect from light, with shaking at 450 rpm.
5. Following 60-minute incubation, wash with Wash Buffer as follows:
• Decant the contents of the wells manually with a hard, rapid downward motion. Fluid should be captured in a receptable designed to collect the waste. Remove all residual reagent from the microplate by tapping it on absorbent paper with the opening facing downwards.
• Fill each well with 300 µL of Washing Buffer with a multichannel pipettor.
• Decant the Washing Buffer from the wells with a hard, rapid downward motion. Remove all residual solution from the microplate by tapping it on absorbent paper with the opening facing downwards.
• Repeat steps ii and iii two more times (total of 3 washings). Do not leave any residual moisture in the wells on each washing step.
6. To each well add 100 µL of Streptavidin-HRP working solution
7. Incubate at room temperature for 20-minutes, covered to protect from light, with shaking at 450 rpm.
8. Following 20-minute incubation, wash with Wash Buffer as follows:
• Decant the contents of the wells manually with a hard, rapid downward motion. Fluid should be captured in a receptable designed to collect the waste. Remove all residual reagent from the microplate by tapping it on absorbent paper with the opening facing downwards.
• Fill each well with 300 µL of Washing Buffer with a multichannel pipettor.
• Decant the Washing Buffer from the wells with a hard, rapid downward motion. Remove all residual solution from the microplate by tapping it on absorbent paper with the opening facing downwards
• Repeat steps ii and iii two more times (total of 3 washings). Do not leave any residual moisture in the wells on each washing step.
9. Next add 100 µL per well of room temperature TMB solution and incubate the plate with TMB solution at room temperature for 20 minutes (covered and protected from light).
10. Then add 100 µL of stop solution per well. Gently tap the plate to mix, ensuring no bubbles are formed, and read plate within 5 minutes after stopping the reaction.
11. On plate reader, measure absorbance at 450 nm with the reference wavelength set at 630–650 nm.

Calcul des résultats:

Follow the steps below to estimate the nuclease concentration of the test samples.

1. Calculate the relative OD 450 using the following formula: Relative OD 450 = (OD 450 of well) – (OD 630-650 nm of the well)
2. Calculate the mean relative OD 450 of the replicates for each standard solution.
3. Plot the standard solutions data as mean relative OD 450 for each standard solution (Y) vs the respective concentration of the standard solutions (X).
4. Fit the standard solution data with a 4-parameter logistic (4-PL) curve. Weight by 1/Y^2 is intended to be used during generation of 4-PL curve
5. Estimate the Nuclease concentration of each test sample well using interpolation from the standard curve. Calculate the average of each respective sample solution concentration.

Note: If the assay samples are from dilutions, multiply the concentrations obtained from interpolations by the dilution factor.
Note: If the spectrometer used for the assay does not automatically subtract the reference wavelength, do this manually.

Précision du teste: Intra-and inter-assay CV% <20%

Restrictions: For Research Use only

Stockage

Stock: 4 °C

Stockage commentaire: The kit and reagents should be stored at 2-8°C. Allow reagents to reach room temperature (18-26°C) before use and may be used until the expiration date. It is recommended to aliquot the reconstituted standard solution and store it at -20°C to avoid freeze/thaw cycles.

Détail du antigène

Antigène: Nuclease

Autre désignation: nucA (Nuclease Produits)

Synonymes: F15D2.37 Kit, F15D2_37 Kit, 5'-3' exonuclease family protein Kit, Nuclease Kit, AT1G29630 Kit, Nuc Kit

Sujet: Nucleases are secreted by Serratia marcescens into the medium it surrounds. The enzyme is a sugar-nonspecific hydrolase, capable of cleaving both RNA and DNA in either double or single stranded form. It requires divalent cations, preferably Mg2+, and is functional across a broad pH range from 6 to 10 (optimal at 8-8.5) and wide temperature ranges between 35°C and 44°C. The ability of Serratia to secrete nuclease appears to be regulated. Bacterial cultures at differing cell densities display different kinetics and efficiencies of nuclease secretion [i.e. growth medium, growth conditions, and host cell mutations]. Anti-Nuclease/NucA Antibody is useful for researcher interested in identifying nucleic acid contamination.