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Ammonium sulfate precipitation is a useful technique as an initial step in protein purification because it enables quick, bulk precipitation of cellular proteins. [4] It is also often employed during the later stages of purification to concentrate protein from dilute solution following procedures such as gel filtration .
Salting out (also known as salt-induced precipitation, salt fractionation, anti-solvent crystallization, precipitation crystallization, or drowning out) [1] is a purification technique that utilizes the reduced solubility of certain molecules in a solution of very high ionic strength.
Calcium carbonate precipitates as a solid, leaving ammonium sulfate in the solution. (NH 4) 2 CO 3 + CaSO 4 → (NH 4) 2 SO 4 + CaCO 3. Ammonium sulfate occurs naturally as the rare mineral mascagnite in volcanic fumaroles and due to coal fires on some dumps. [14] Ammonium sulfate is a byproduct in the production of methyl methacrylate. [15]
Protein purification is a series of processes intended to ... ammonium sulfate can be ... these ligands function in a fashion similar to that of antibody-antigen ...
The ideal salt for protein precipitation is most effective for a particular amino acid composition, inexpensive, non-buffering, and non-polluting. The most commonly used salt is ammonium sulfate. There is a low variation in salting out over temperatures 0 °C to 30 °C.
Pure results of a successful purification process are termed isolate. The following list of chemical purification methods should not be considered exhaustive. Affinity purification purifies proteins by retaining them on a column through their affinity to antibodies, enzymes, or receptors that have been immobilised on the column.
Immunoprecipitation of intact protein complexes (i.e. antigen along with any proteins or ligands that are bound to it) is known as co-immunoprecipitation (Co-IP). Co-IP works by selecting an antibody that targets a known protein that is believed to be a member of a larger complex of proteins.
KLH purification may also include endotoxin removal, but this step is often unnecessary because the endotoxin serves as an adjuvant when injected for antibody production. If the protein becomes denatured or if the copper ions are lost in the purification process, the opalescent blue color disappears and the solution becomes a dull grayish color.