Article: Hapten-conjugated antibodies and visual markers used to label cell-surface antigens for electron microscopy: an approach to double labeling

Haptens can be coupled to mouse immunoglobulin with retention of antibody specificity. Such hapten-coupled alloantibody, when bound to cells, can be bridged to an electron microscopic marker in two ways: (i) by a hybrid antibody, i.e., anti-hapten:anti-marker, and (ii) by untreated bivalent anti-hapten antibody if the marker is also haptenized. Through the use of immunologically non-crossreactive haptens and markers it should be possible to localize multiple, different antigens on the surface of an individual cell. Double labeling has been achieved with the first method.

Citing Articles

Hapten-sandwich labeling. I. A general procedure for simultaneous labeling of multiple cell surface antigens for fluorescence and electron microscopy.

Wolfsy L, Baker P, Thompson K, Goodman J, Kimura J, Henry C J Exp Med. 1974; 140(2):523-37.

PMID: 4602983 PMC: 2139603. DOI: 10.1084/jem.140.2.523.

Hapten-sandwich labeling. II. Immunospecific attachment of cell surface markers suitable for scanning electron microscopy.

Nemanic M, Carter D, Pitelka D, WOFSY L J Cell Biol. 1975; 64(2):311-21.

PMID: 163829 PMC: 2109504. DOI: 10.1083/jcb.64.2.311.

References

1.

Farquhar M, Palade G . Cell junctions in amphibian skin. J Cell Biol. 1965; 26(1):263-91. PMC: 2106698. DOI: 10.1083/jcb.26.1.263. View

2.

Brownstone A, Mitchison N, PITT-RIVERS R . Chemical and serological studies with an iodine-containing synthetic immunological determinant 4-hydroxy-3-iodo-5-nitrophenylacetic acid (NIP) and related compounds. Immunology. 1966; 10(5):465-79. PMC: 1423692. View

3.

Hammerling U, Aoki T, de Harven E, Boyse E, Old L . Use of hybrid antibody with anti-gamma-G and anti-ferritin specificities in locating cell surface antigens by electron microscopy. J Exp Med. 1968; 128(6):1461-73. PMC: 2138585. DOI: 10.1084/jem.128.6.1461. View

4.

LEVINE B, Levytska V . Hapten-specific insoluble immunoabsorbents prepared from hide powder (collagen). J Immunol. 1969; 102(3):647-55. View

5.

Hammerling U, Aoki T, Wood H, Old L, Boyse E, de Harvin E . New visual markers of antibody for electron microscopy. Nature. 1969; 223(5211):1158-9. DOI: 10.1038/2231158a0. View

6.

Silvestre D, KOURILSKY F, Nicolai M, LEVY J . Presence of HLA antigens on human reticulocytes as demonstrated by electron microscopy. Nature. 1970; 228(5266):67-8. DOI: 10.1038/228067a0. View

7.

Frye L, Edidin M . The rapid intermixing of cell surface antigens after formation of mouse-human heterokaryons. J Cell Sci. 1970; 7(2):319-35. DOI: 10.1242/jcs.7.2.319. View

8.

Nicolson G, HYMAN R, Singer S . The two-dimensional topographic distribution of H-2 histocompatibility alloantigens on mouse red blood cell membranes. J Cell Biol. 1971; 50(3):905-10. PMC: 2108302. DOI: 10.1083/jcb.50.3.905. View

9.

Aoki T, Wood H, Old L, Boyse E, de Harven E, LARDIS M . Another visual marker of antibody for electron microscopy. Virology. 1971; 45(3):858-62. DOI: 10.1016/0042-6822(71)90214-5. View

10.

Nisonoff A, RIVERS M . Recombination of a mixture of univalent antibody fragments of different specificity. Arch Biochem Biophys. 1961; 93:460-2. DOI: 10.1016/0003-9861(61)90296-x. View

11.

Reynolds E . The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963; 17:208-12. PMC: 2106263. DOI: 10.1083/jcb.17.1.208. View

12.

Nisonoff A . ENZYMATIC DIGESTION OF RABBIT GAMMA GLOBULIN AND ANTIBODY AND CHROMATOGRAPHY OF DIGESTION PRODUCTS. Methods Med Res. 1964; 10:134-41. View

Hapten-conjugated Antibodies and Visual Markers Used to Label Cell-surface Antigens for Electron Microscopy: an Approach to Double Labeling