Certolizumab Pegol specific ELISA 试剂盒

• 1 x 12 x 8 Microtiter Plate Break apart strips coated with anti-Cerolizumab pegol monoclonal antibody.
• 5 x 0.3 mL Certolizumab pegol Standards A-E, Concentrate (500x), 100, 30, 10, 3, and 0 µg/mL. Used for construction of the standard curve. Contains Certolizumab pegol, proteins, stabilizer and <15mM NaN3.
• 1 x 12 mL Assay Buffer Blue colored. Ready to use. Contains proteins and <15mM NaN3.
• 1 x 60 mL Dilution Buffer, Concentrate (5X), Contains proteins and <15mM NaN3.
• 1 x 12 mL Biotinylated TNFα Green colored. Ready to use. Contains biotinylated recombinant human tumor necrosis factor alpha (rhTNFα), proteins, stabilizers and <15 mM NaN3.
• 1 x 12 mL Enzyme Conjugate Red colored. Ready to use. Contains horseradish peroxidase(HRP)-conjugated streptavidin (HRP-Streptavidin), Proclin® and stabilizers.
• 1 x 12 mL TMB Substrate Solution Ready to use. Contains 3,3',5,5'-Tetramethylbenzidine (TMB).
• 1 x 12 mL Stop Solution Ready to use. 1 N Hydrochloric acid (HCl).
• 1 x 50 mL Wash Buffer, Concentrate (20x) Contains buffer, Tween® 20 and KathonTM.
• 2 x 1 Adhesive Seal For sealing microtiter plate during incubation.

• Micropipettes (< 3 % CV) and tips to deliver 5-1000 μL.
• Bidistilled or deionised water and calibrated glasswares (e.g. flasks or cylinders).
• Wash bottle, automated or semi-automated microtiter plate washing system.
• Microtiter plate reader capable of reading absorbance at 450 nm (reference wavelength at 600-650 nm is optional).
• Absorbent paper towels, standard laboratory glass or plastic vials, and a timer.

• Before performing the assay, samples and assay kit should be brought to room temperature (about 30 minutes beforehand) and ensure the homogeneity of the solution.
• All Standards should be run with each series of unknown samples.
• Standards should be subject to the same manipulations and incubation times as the samples being tested.
• All steps of the test should be completed without interruption.
• Use new disposable plastic pipette tips for each reagent, standard or specimen in order to avoid cross contamination.

ELISA Kits are suitable also for using by an automated ELISA processor.

Dilution of Standards and Samples
The dilutions depicted below are examples of how to obtain final 1:500 dilution. Standards and samples should be properly diluted as homogenous mixture before starting the assay procedure. It is recommended mixing the standards and samples well to homogenous solution at each dilution step.
1. 10 µL of standard or sample added to 90 µL of 1X dilution buffer, giving the total volume of 100 µL and a dilution of 1:10.
2. 10 µL of 1:10 diluted standard or sample added to 490 µL of 1X dilution buffer, giving the total volume of 500 µL and a dilution of 1:500.
3. Samples with a drug concentration above the measuring range should be rated as “>highest standard”. The result should not be extrapolated. The sample in question should be further diluted with 1X Dilution Buffer and then retested.

A standard curve should be calculated using the standard concentration (X-axis) versus the OD450 (or OD450/620) values (Y-axis). This can be done manually using graph paper or with a computer program. Concerning the data regression by computer, it is recommended to primarily use the “4 Parameter Logistic (4PL)” or alternatively the “point-to-point calculation”. In case of manual plot there are 2 options: Semilog graph or linear graph. The concentration of the samples can be read from this standard curve as follows. Using the absorbance value for each sample, determine the corresponding concentration of the drug from the standard curve. This value always has to be multiplied by the individual dilution factor, which usually will be 500. If any diluted sample is reading greater than the highest standard, it should be further diluted appropriately with 1X Dilution Buffer and retested. Also this second dilution has to be used for calculation the final result.