The unconjugated immunoglobulin gives a single arc when tested by immunoelectrophoresis and 2D-IEPagainst human serum.
Identity has been confirmed by double diffusion (Ouchterlony) against human serum and an anti-humantransferrin of known specificity.
The immunogen used was Human Holo-Transferrin purified from human serum and shown to be homogeneousby SDS-PAGE.
Shipped at 4°C. Store at +4°C.
PBS pH7.2,ProclinTM300 (0.05%)
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Antiserum is prepared by immunisation of sheep with the above immunogen and if necessary, adsorption to monospecificity by the use of solid phase adsorbants.
An immunoglobulin fraction is thenproduced by ion-exchange chromatography and zone electrophoresis is performed to ensure that only agamma-mobility component is present.
The antibody is then conjugated with horseradish peroxidase activated by periodate.
Preservatives areadded and the product is then 0.2µm filtered.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
For enzyme immunoassays, to give an A450nm = 1.0 using wells coated with transferrin at 2µg/mLand TMB as a substrate: 1/12800 – 1/25600
The titres reported should be used as guidelines for determining the working dilution in the userssystem.
The following points should be observed when using this product:
1) Dilutions of the conjugate must be made in a suitable buffer (e.g. 0.15M Phosphate buffered saline,pH7.2)
2) It is recommended that dilutions of the conjugate are made in polypropylene or polyethylene containers.Other materials (e.g. polystyrene, some grades of glass) may be detrimental to the conjugate.
3) Working dilutions of the conjugate should be prepared immediately prior to use.
4) Peroxidase conjugates are adversely affected by the presence of sodium azide, the use of which shouldtherefore be avoided.
5) The addition of Tween 20 (0.05 – 0.1%) to the conjugate diluent buffer is recommended to prevent nonspecificbinding in EIA. It may also be advantageous in other assay systems.
Transferrins are iron binding transport proteins which can bind two Fe(3+) ions in association with the binding of an anion, usually bicarbonate. It is responsible for the transport of iron from sites of absorption and heme degradation to those of storage and utilization. Serum transferrin may also have a further role in stimulating cell proliferation.
Expressed by the liver and secreted in plasma.
Defects in TF are the cause of atransferrinemia (ATRAF) [MIM:209300]. Atransferrinemia is rare autosomal recessive disorder characterized by iron overload and hypochromic anemia.
Belongs to the transferrin family. Contains 2 transferrin-like domains.