(Myoglobin, BSA or Cytochrome C) using an ion-exchange chromatography method via a cation (CM) or anion (DEAE) exchanger, followed by a SDS-PAGE gel electrophoresis technique to find the concentration of the sample proteins, with the help of a Bradford assay, which employs a Coomassie dye to bind to proteins. After all techniques were performed, Protein 1 was found to have an absorbance value of 0.451 and a concentration of 0.019 µg/µL. Protein 2 was found to have an absorbance of 0.373 and a concentration
with IL-1β and production of monoclonal antibodies, iv) Establishment of a sandwich ELISA (Enzyme Linked Immunosorbent Assay) with the antibodies and the antigen. These steps were successfully completed recombinant IL-1β was produced in a reliable quality and sufficient quantity. Monoclonal (mice) and polyclonal (sheep) antibodies against rabbit IL-1β were isolated. The ELISA assay allows the quantitative determination of the endogenous rabbit fever signal IL-1β. Comparison of three pyrogen tests
Research Methodology Microorganism Newly isolated Aspergillus niger that was provided by Dr. Bilqees Fatima, Assistant Professor Botany Department Minhaj University Lahore. Substrates Seven different kinds of agro-industrials substrates was used such as wheat bran, wheat straw, rice straw, sugar cane, brassica napus, gram testa and oat. All of these substrates was brought from local market of Lahore. Ten gram of each substrate was used in this study and 10ml of distilled water was added in it.
(TCA) was added and centrifuged at 845 x g for 15 min at 4◦C. Further, 0.5 ml of the supernatant was added to 2.5 ml of 0.01% DTNB and the absorbance was read at 412 nm (Figure 2.19). The protein present in the pellet was determined by the Bradford method (Bradford, 1976) and the concentration of GSH was expressed as mmol/mg protein. Different concentrations of glutathione were taken as standard and data was represented as μmoles/mg protein.