An assay for intrinsic factor based on blocking of the R binder of gastric juice by cobinamide.
نویسندگان
چکیده
An in vitro assay for measurement of gastric juice intrinsic factor (IF) was developed based on the ability of the cobinamide (Cbi) [(CN, OH) Cbi] to bind to the gastric juice R-type binders of cobalamin (Cbl) and not to the IF binder. Subsequently added radioactive Cbl, CN-[57Co] Cbl, binds only to the IF binders and allows for direct measurement of this Cbl binding protein. This Cbi blocking assay was found to function as well as the more conventional methods of IF measurement, G-100 column chromatography, and IF blocking antibody assay. The present assay has the advantage of eliminating the need for elaborate forms of protein separation and does not rely on a source of antibody.
منابع مشابه
Commercially-Supplied Binders for Plasma Cobalamin (Vitamin B12), Analysis-’ ‘Punt led” Intrinsic Factor, “Cobinamide’ ‘-Blocked R-Protein Binder, and Non-purified Intrinsic Factor-R-Protein Binder-Compared to Microbiological Assay
Recent evidence (Kolhouse et aL, N. EngI. J. Med. 299: 785-792, 1978) demonstrates that commercial cobalamin (Vitamin B12) radioassay kits contain nonspecific R-protein bindingagents thatcan give falsely normal results in patients who are actually cobalamin deficient. We tested three kits:with“purified” intrinsic factor as the binder, with intrinsic factor and the nonspecific R-protein sites bl...
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The binding of vitamin B(12) by human gastric juice has been found to be pH dependent. Maximum binding occurs between pH 6.5 and 10. Outside this pH range the vitamin B(12)-binding ability of human gastric juice decreases and at pH below 2 or above 12.2 this drops sharply to about 10 to 15% of the maximum. Three commercial hog intrinsic factors have been found to give a similar response to pH c...
متن کاملPreparation and characterization of proteins in the alimentary tract of the dog which bind cobalamin and intrinsic factor.
The pyloric mucosa and the gastric and intestinal juices of the dog were found to contain two cobalaminbinding proteins which have been identified as intrinsic factor and R binder. T h e ileal mucosa also contains unsaturated R binder and, in addition, a specific receptor protein which couples the intrinsic factormcobalamin complex. This receptor protein was solubilized using mechanical sharing...
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Intrinsic factor can bind vitamin B12 and simultaneously attach to receptors on liver or small intestine in a calcium-dependent manner (1). There appears to be a stoichiometric relationship between the receptors, intrinsic factor and vitamin B12 (2). This report, previously presented in preliminary form (3), is concerned with blocking of the receptor sites for hog intrinsic factor by human gast...
متن کاملBlocking of Hog Intrinsic Factor by Human Gastric Juice and Certain Mucopolysaccharides , in - Cluding Blood
Intrinsic factor can bind vitamin B12 and simultaneously attach to receptors on liver or small intestine in a calcium-dependent manner (1). There appears to be a stoichiometric relationship between the receptors, intrinsic factor and vitamin B12 (2). This report, previously presented in preliminary form (3), is concerned with blocking of the receptor sites for hog intrinsic factor by human gast...
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ورودعنوان ژورنال:
- Blood
دوره 53 4 شماره
صفحات -
تاریخ انتشار 1979