Literature DB >> 4505654

Solid-phase conjugation of ferritin to Fab-fragments of immunoglobulin G for use in antigen localization on thin sections.

J P Kraehenbuhl, J D Jamieson.   

Abstract

A technique for the solid-phase conjugation of ferritin to monovalent antibodies (Fab-fragments of immunoglobulin G) is described in which Fab-fragments from sheep antiserum to rabbit Fab-fragments are first adsorbed to Sepharose containing insolubilized rabbit immunoglobulin G, and subsequently allowed to couple with ferritin previously activated with toluene-2,4-diisocyanate. The conjugate, after elution from the adsorbent, is separated from free ferritin and unconjugated Fab by gel chromatography. We have used this conjugate as a common second-step reagent for the surface localization of intracellular antigens on thin sections of tissue embedded in albumin. The first-step reagent consists of specific rabbit antibodies (IgG) directed against the tissue component under study. An example of the use of this technique for the localization of trypsinogen in bovine exocrine pancreatic cells is given.

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Year:  1972        PMID： 4505654      PMCID： PMC426799          DOI： 10.1073/pnas.69.7.1771

Source DB:  PubMed          Journal:  Proc Natl Acad Sci U S A        ISSN： 0027-8424            Impact factor:   11.205


  14 in total

1.  On the formation of covalent linkages between two protein molecules.

Authors:  A F SCHICK; S J SINGER
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2.  Enzyme-labeled antibodies for the light and electron microscopic localization of tissue antigens.

Authors:  P K Nakane; G B Pierce
Journal:  J Cell Biol       Date:  1967-05       Impact factor: 10.539

3.  A general method for the specific staining of intracellular antigens with ferritin-antibody conjugates.

Authors:  J D McLean; S J Singer
Journal:  Proc Natl Acad Sci U S A       Date:  1970-01       Impact factor: 11.205

4.  Ultrastructural localization of intracellular immune globulins in plasma cells and lymphoblasts by enzyme-labeled antibodies.

Authors:  E H Leduc; G B Scott; S Avrameas
Journal:  J Histochem Cytochem       Date:  1969-04       Impact factor: 2.479

5.  Immunochemical quantitation of antigens by single radial immunodiffusion.

Authors:  G Mancini; A O Carbonara; J F Heremans
Journal:  Immunochemistry       Date:  1965-09

6.  Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads.

Authors:  P Cuatrecasas
Journal:  J Biol Chem       Date:  1970-06       Impact factor: 5.157

7.  Localization of tissue antigens on the ultrathin sections with peroxidase-labeled antibody method.

Authors:  Y Kawarai; P K Nakane
Journal:  J Histochem Cytochem       Date:  1970-03       Impact factor: 2.479

8.  Ultrastructural localization of intracellular antigen using enzyme-labeled antibody fragments.

Authors:  J P Kraehenbuhl; P B De Grandi; M A Campiche
Journal:  J Cell Biol       Date:  1971-08       Impact factor: 10.539

9.  Intracellular transport of secretory proteins in the pancreatic exocrine cell. I. Role of the peripheral elements of the Golgi complex.

Authors:  J D Jamieson; G E Palade
Journal:  J Cell Biol       Date:  1967-08       Impact factor: 10.539

10.  The localization of spectrin on the inner surface of human red blood cell membranes by ferritin-conjugated antibodies.

Authors:  G L Nicolson; V T Marchesi; S J Singer
Journal:  J Cell Biol       Date:  1971-10       Impact factor: 10.539
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  19 in total

1.  Ferritin-conjugated antibodies used for labeling of organelles involved in the cellular synthesis and transport of procollagen.

Authors:  B R Olsen; D J Prockop
Journal:  Proc Natl Acad Sci U S A       Date:  1974-05       Impact factor: 11.205

2.  Attempts to quantitate immunocytochemistry at the electron microscope level.

Authors:  J P Kraehenbuhl; L Racine; G W Griffiths
Journal:  Histochem J       Date:  1980-05

3.  Synthesis and intracellular transport of proteins in the exocrine pancreas of the frog (Rana esculenta). I. An ultrastructural and autoradiographic study.

Authors:  J W Slot; J J Geuze; C Poort
Journal:  Cell Tissue Res       Date:  1974       Impact factor: 5.249

4.  Immunochemistry on ultrathin frozen sections.

Authors:  K T Tokuyasu
Journal:  Histochem J       Date:  1980-07

Review 5.  A review of macromolecular transport and secretion at the cellular level.

Authors:  M Singh; P J Webster
Journal:  Am J Dig Dis       Date:  1976-04

6.  Use of the avidin-biotin complex for specific staining of biological membranes in electron microscopy.

Authors:  H Heitzmann; F M Richards
Journal:  Proc Natl Acad Sci U S A       Date:  1974-09       Impact factor: 11.205

7.  Immunoferritin localization of intracellular antigens: the use of ultracryotomy to obtain ultrathin sections suitable for direct immunoferritin staining.

Authors:  R G Painter; K T Tokuyasu; S J Singer
Journal:  Proc Natl Acad Sci U S A       Date:  1973-06       Impact factor: 11.205

8.  Ultrastructural localization of sucrases in Streptococcus mutans GS-5 and an extracellular polysaccharide mutant: a comparative cytochemical and immunocytochemical study.

Authors:  J J Bozzola; M C Johnson; I L Shechmeister
Journal:  Infect Immun       Date:  1977-08       Impact factor: 3.441

9.  Localization of acetylcholine receptor in excitable membrane from the electric organ of Torpedo: Evidence for exposure of receptor antigenic sites on both sides of the membrane.

Authors:  R Tarrab-Hazdai; B Geiger; S Fuchs; A Amsterdam
Journal:  Proc Natl Acad Sci U S A       Date:  1978-05       Impact factor: 11.205

10.  Location of procollagen in chick corneal and tendon fibroblasts with ferritin-conjugated antibodies.

Authors:  C Nist; K Von Der Mark; E D Hay; B R Olsen; P Bornstein; R Ross; P Dehm
Journal:  J Cell Biol       Date:  1975-04       Impact factor: 10.539
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