Trastuzumab Kit ELISA

N° du produit ABIN4886397

Détail du produit Trastuzumab Kit ELISA

Antigène: Trastuzumab

Reactivité: Humain, Souris, Rat

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 1.25-40 ng/mL

Seuil minimal de détection: 1.25 ng/mL

Application: ELISA

Fonction: Quantification of Trastuzumab in biological matrices

Type d'échantillon: Plasma, Serum

Analytical Method: Quantitative

Specificité: Trastuzumab (Herceptin)

Réactivité croisée (Details): hIgG1, Rituximab, and Infliximab prepared at 250 ng/mL were assayed and exhibited no cross-reactivity or interference.

Ingrédients: Coated microtiter plate, 96 wells
Calibrator diluent. - 1.8ml
Calibrator 12ul
10X wash buffer - 25ml
Assay buffer - 50ml
1000X detection reagent - 17ul
TMB - 12ml
TMB stop solution - 12ml
Plate sealers - 3

Matériel non inclus: Precision pipettes calibrated to deliver 5-1000μL
Multi-channel pipette calibrated to deliver 50-200μL
Plate shaker
Disposable tips
Vortex-Mixer
Distilled or de-ionized water
Microplate reader capable of reading 450nm with background subtrac

Information d'application

Indications d'application: Optimal working dilution should be determined by the investigator.

Volume d'échantillon: 15 μL

Durée du test: 2.5 h

Plaque: Pre-coated

Protocole: The Trastuzumab ELISA kit is designed to measure free Trastuzumab with high specificity and sensitivity . This assay employs the sandwich enzyme immunoassay technique. A precoated anti-Trastuzumab 96 well plate is provided. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Trastuzumab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, enzyme linked detection antibody is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of Trastuzumab present in test samples and the concentration is calculated from the standard series.

Préparation des réactifs:

Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label. 1. Wash Buffer (1X) Preparation: Dilute wash buffer concentrate with ultra-pure water 1/10 before use (for example add 10 mL concentrate to 90 mL ultra-pure water). Mix well. 2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 10 μL concentrate to 10 mL of assay buffer). Mix welll. 3. Preparation of Calibrators: Prepare calibrators with concentrations ranging from 1000 ng/mL to 62.5 ng/mL. The following is an example calibrator curve.

Prélèvement de l'échantillon: This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20 °C for up to 1 year.

Préparation de l'échantillon:

Dilute calibrators and test samples 1/50 with assay buffer (for example add 5μL of prepared calibrator or sample to 245μL of assay buffer). Mix well. Do not store diluted samples.

Procédure de l'essai:

This assay employs the sandwich enzyme immunoassay technique. Anti- Trastuzumab is coated onto a 96 well microplate. Calibrator, quality control samples (if desired) and test samples are pipetted into the appropriate wells. Trastuzumab present in biological matrices is bound by the immobilized anti- Trastuzumab antibody. After washing away any unbound substances, enzyme linked antiTrastuzumab antibody is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of Trastuzumab present in test samples. The color development is stopped and the intensity of the color is measured.

Calcul des résultats:

1. Construct a standard curve by plotting the absorbance obtained from each standard against concentration. Use a 4 or 5 parameter curve fit. Alternatively a log-log curve fit may be used. 2. The concentration of the unknowns can be read directly from this standard curve using the absorbance value for each sample. 3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with calibrator diluent and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.

Précision du teste: Precision: The precision was determined by analyzing samples prepared at 1000 ng/mL in 6 replicates on 6 different occasions. Intra-assay coefficient of variation (CV) < 10%. Inter-assay CV < 10%.

Recovery: 1000ng/mL of Trastuzumab was spiked in 10 lots of human serum. Recovery ranges are from 91-117% with an average recovery of 106%.

Restrictions: For Research Use only

Stockage

Agent conservateur: Without preservative

Précaution d'utilisation: Read manual completely before beginning

Stock: -20 °C

Stockage commentaire: Store kit components at -20°C unless specified otherwise. DO NOT USE past kit expiration date. Some vials contain a small amount of reagents. Spin tubes on pulse setting prior to opening.

Date de péremption: 12 months

Détail du antigène

Antigène: Trastuzumab

Abstract: Trastuzumab Produits

Sujet: Trastuzumab (Herceptin®) is indicated for the treatment of HER2-positive breast cancer, and adjuvant therapies for metastatic gastric cancer and gastroesophageal cancer. Serum concentration of Trastuzumab may predict some clinical outcome during therapy. It is also possible that the surveillance of circulating Trastuzumab concentration during therapy represents a direct factor for immunogenicity and some other side effects. Identification of biomarkers and risk factors for adverse drug reactions that might be related to serum concentrations, and maintaining the effective minimum concentration of Trastuzumab in order to potentially avoid some side effects, might be beneficial using a reliable method

ID gène: 2064