Labeling of proteins with 125I and experimental determination of its specific activity

Caro, R.A.; Ciscato, V.A.; Giacomini, S.M.V. De; Quiroga, Santiago; Radicella, R.

doi:10.1016/0020-708x(75)90091-5

Cited by 17 publications

(2 citation statements)

References 7 publications

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“…In the general case of macromolecules, the degree of incorporation (fraction of bound iodine versus fraction of free iodine) is measured on a sample aliquot by chromatography electrophoresis (CARO et al 1975) or protein precipitation by trichloroacetic acid). In the case of small molecules (haptens), it is generally possible to purify the different molecular species and identify them .…”

Section: Specific Radioactivitymentioning

confidence: 99%

Radioiodination and Other Labeling Techniques

Grassi¹,

Maclouf²,

Pradelles³

1987

Radioimmunoassay in Basic and Clinical Pharmacology

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Section: Specific Radioactivitymentioning

confidence: 99%

Radioiodination and Other Labeling Techniques

Grassi¹,

Maclouf²,

Pradelles³

1987

Radioimmunoassay in Basic and Clinical Pharmacology

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“…Iodination of IgG and factor VIII was achieved using the chloramine-T method, modified and adapted from the methods of Hallaba and Drouet (21) and Caro et al (22). For IgG, 50 pl protein solution (4.5-5.0 mglml) were used, to which was added sufficient labelling buffer 1O.bS fvf phosphate, pH 7.4), followed by 500 pCi 12s1 o, 1,200 p,gi r3r1 to gir" u volume of 720 pl.…”

Section: Iodine-labelled Factor Viil and Isgmentioning

confidence: 99%

Studies on Human Factor VIII and its Antibodies Using Radiolabelling and Affinity Chromatography

1981

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SummaryAn immuno-affinity chromatography method was used to isolate human factor VIII and its antibodies and the mechanism of the affinity system was investigated using iodine labelling.Rabbit antibodies to human factor VIII were insolubilised onto CNBr — activated Sepharose 2B which was used for the preparation of affinity columns. Both VIII:C and VIIIR:Ag were adsorbed onto such columns from factor VIII preparations. The subsequent application of immunoglobulin preparations containing human antibodies to factor VIII resulted in the adsorption of these antibodies onto the columns. Adsorbed material was eluted from the affinity columns with 0.2 M glycine - HCl, pH 2.3.When 125I-labelled factor VIII and 131I-labelled human antibodies to factor VIII were used in this affinity system, the eluted material could be separated into three fractions by gel filtration on Bio-Gel A 1.5 m. Fraction 1 occurred at the void volume position, fraction 3 at a position corresponding to the elution position of IgG and fraction 2 at an intermediate position. 131I-labelled material was present in all three peaks. 125I-labelled material was present mainly in peak 1, with a little in peak 2. The results support the view that VIIIR: Ag, which binds heterologous antibodies, is non-covalently linked to a smaller subunit, VIII.C, which binds homologous antibodies.

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A comparative study of radioiodination of simple aromatic compounds via N‐halosuccinimides and chloramine‐t in TFAA

Youfeng

Coenen

Petzold

et al. 1982

Labelled Comp Radiopharmac

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e t z o l d a n d G . S t o c k l i n I n s t i t u t f u r Chemie 1 ( N u k l e a r c h e m i e ) , K e r n f o r s c h u n g s a n l a g e J u l i c h G m b H , D-5170 J u l i c h , FRG SUMMARY A c o m p a r a t i v e s t u d y o f r a d i o i o d i n a t i o n o f s i m p l e a r o m a t i c compounds u s i n g N -c h l o r o s u c c i n i m i d e (NCS), N -b r o m o s u c c i n i m i d e (NBS), N -c h l o r o t e t r a f l u o r os u c c i n i m i d e (NCTFS) a n d Chloramine-T (CAT) h a s b e e n c a r r i e d o u t i n t r i f l u o r o a c e t i c a n h y d r i d e (TFA?.) a s a n a p r o t i c s o l v e n t . O p t i m i z a t i o n of t h e i o d i n a t i o n p r o c e d u r e h a s b e e n p e r f o r m e d u s i n g a n i s o l e a s t h e model s u b s t r a t e . A f a s t and s i m p l e " o n e -p o t ' ' s y n t h e s i s v i a NCTFS, NCS o r CAT l e a d s t o i o d o a n i s o l e s w i t h a r a d i o c h e m i c a l y i e l d of a b o u t 7 0 8 . F o r c o m p a r i s o n , t o l u e n e a n d b e n z e n e h a v e a l s o b e e n r a d i o i o d i n a t e d . Key w o r d s : I o d i n a t i o n , I o d i n e -I 2 3 , 1 2 5 , 1 3 1 , N -h a l o s u c c i n i m i d e , Chloramine-T.

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Labeling of proteins with 125I and experimental determination of its specific activity

Cited by 17 publications

References 7 publications

Radioiodination and Other Labeling Techniques

Radioiodination and Other Labeling Techniques

Studies on Human Factor VIII and its Antibodies Using Radiolabelling and Affinity Chromatography

A comparative study of radioiodination of simple aromatic compounds via N‐halosuccinimides and chloramine‐t in TFAA

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