AffinityImmuno

ELISA/assay

Adalimumab (Humira) Pharmacokinetic ELISA

EL-1611-011

1X96 wells

599.99 USD

EL-1611-011

Adalimumab (Humira) Pharmacokinetic ELISA

ELISA

599.99 USD

479.99

USD

1X96 wells

-20C, 1 year

Adalimumab (Humira)

Human, mouse, rat

50ng/ml - 1.56ng/ml

1.5ng/ml

The Adalimumab ELISA kit is designed to measure free Adalimumab with high specificity and sensitivity. This assay employs the sandwich enzyme immunoassay technique. A precoated anti-Adalimumab 96 well plate is provided. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Adalimumab 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 Adaliumumab present in test samples and the concentration is calculated from the standard series.

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

Direct sandwich ELISA

Peroxidase / OD450

Quantification of Adalimumab in biological matrices

Strip

Serum Plasma

15ul

2.5 hours

Adalimumab

<10%, <10%

Adalimumab (Trade name Humira®) is a human monoclonal antibody used to block the action of tumor necrosis factor alpha (TNF- ). TNF-α is a strong proinflammatory cytokine involved in normal immunity and also various autoimmune diseases. The Adalimumab ELISA kit is designed to measure free Adalimumab with high specificity and sensitivity. The assay design uses a pair of antibodies allowing detection of whole Adalimumab molecules in biological matrices

This assay employs the sandwich enzyme immunoassay technique. Anti- Adalimumab is coated onto a 96 well microplate. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Adalimumab 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 Adalimumab present in test samples.

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 20mL concentrate to 180mL ultra-pure water). Mix well. 2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 11μl concentrate to 11ml of assay buffer). Mix well. 3. Preparation of Calibrators: Prepare calibrators with concentrations ranging from 2,500 ng/mL to 78 ng/mL. The following is an example calibrator curve.

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.

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.

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.