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Golimumab ELISA is suitable also for using by an automated ELISA processor.
Wash Buffer: Dilute 10 mL Wash Buffer (up to 200 mL) at the ratio of 1:20 with distilled water.Warm up at 37 °C to dissolve crystals. Mix vigorously.Store at 2-8 °C for up to 4 weeks.Prepare Wash Buffer before starting the assay procedure.
Serum/ Plasma: Initially dilute the Serum/ Plasma (Sample) at the ratio of 1:20 with Assay Buffer.Sample : Assay Buffer Relation can be 1:20-1:100.For dilution at 1:20, 10 μL Sample + 190 μL Assay BufferFor dilution at 1:100, 5 μL Sample + 495 μL Assay Buffer If any sample, initially diluted as indicated above, produces an OD value above the measuring range it should be rated as "> highest standard". The result should not be extrapolated. The sample in question should be further diluted with Assay Buffer and then retested.Serum, Plasma (EDTA, Heparin): The usual precautions for venipuncture should be observed. It is important to preserve the chemical integrity of a blood specimen from the moment it is collected until it is assayed. Do not use grossly hemolytic, icteric or grossly lipemic specimens. Samples appearing turbid should be centrifuged before testing to remove any particulate material. Storage: 2-8 °C &leq,-20 °C (Aliquots)Keep away from heat or direct sun light.Avoid repeated freeze-thaw cycles.Stability: 3 days at 2-8 °C, 6 months at -20 °C
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 we are recommending 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 . Semilog graph paper is available at http://www.papersnake.com/logarithmic/semilogarithmic/. 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 dilution factor. If any diluted sample is reading greater than the highest standard, it should be further diluted appropriately with Assay Buffer and retested. Also this second dilution has to be used for calculation the final result.