Protein L-Ligand Leakage Elisa

This ELISA-kit has been designed to detect and quantify Protein L in Immunoglobulin (Ig) or Ig-fragment containing solutions. It is a sandwich ELISA with microtiter strips coated with an affinity purified anti-Protein L IgY-antibody. In this assay, samples are boiled to separate the Ig/Ig-fragments from the Protein L in the solution, centrifuged, and incubated in the strip wells. The bound Protein L is then detected by adding a horseradish peroxidase (HRP)-conjugated anti-Protein L IgY antibody. Substrate is added to induce a color change which is proportional to the amount of bound Protein L from the sample. The color change is quantified spectrophotometrically at 450nm by using a plate reader. The amount of protein L in the sample is determined by using a standard curve prepared in a reference sample. The reference sample should have the same concentration of Ig or Ig-fragments and preferably the same buffer composition as the elution samples from the Protein L affinity chromatography medium. However, it should be prepared using another purification strategy, e.g. ion exchange chromatography. Once a reference sample is created it can be aliquoted and used for several different protein L ELISA assays. Applications: Determination of leakage levels of protein L in protein L-medium purification of intact immunoglobulin or immunoglobulin fragments (Ig or Ig fragments). Some fab samples may require optimization. Currently there is no protocol available for sdab molecules. Shipping and storage. The product is shipped cooled (+4°C) except the protein L reference which is shipped frozen (-20°C). On arrival store components cooled (+4ºC) except the protein L reference which should be stored at -20/-80°C: Strips (96 wells), store unused strips in sealed bag at +4ºC. HRP-conjugated anti Protein L pAb and EC-Blue Enhanced TMB-substrate, store protected from light. Protein L reference,store in working aliquots to avoid repeated freezing/thawing.