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PMID: 6811596 Published · ppublish English Journal Article

Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line.

The Journal of cell biology ·Vol. 93 ·No. 3 ·1982-06-00 ·Pages 576-82

Kilmartin JV, Wright B, Milstein C

Abstract

Hybrid myeloma cell lines secreting monoclonal antibodies to tubulin have been prepared using rat myelomas and spleen cells from rats immunized with yeast tubulin. A comparison between the results obtained with the rat myeloma Y3-Ag 1.2.3., which secretes a light chain, and a new line, YB2/O, which does not, shows that they are both excellent parental lines and that the second produces hybrids with no myeloma chain components. The antitubulin antibodies in the serum of rats bearing two of the hybrid myeloma tumors gave titers of up to 1:10(6) from which large amounts of monoclonal antibodies could be easily purified. They recognized tubulin from yeast as well as from birds and mammals. The two antibodies gave clear immunofluorescent staining of yeast mitotic spindles as well as the interphase microtubule network of tissue culture cells. Some difference in the pattern of immunofluorescence staining of yeast cells and nuclei was observed between the two antibodies. The purified antibodies could be conjugated to colloidal gold particles and used for direct labeling of yeast microtubules for electron microscopy.

MeSH Terms
Animals Antibodies, Monoclonal/biosynthesis,immunology Cell Line Fluorescent Antibody Technique Hybrid Cells Immunoglobulin Light Chains/biosynthesis Melanoma Microtubules/immunology Rats Tubulin/immunology Yeasts
Chemicals
Antibodies, Monoclonal Immunoglobulin Light Chains Tubulin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kilmartin J V
Wright B
Milstein C
References (33)
33 references, click to expand
  1. Meiotic spindle plaques in Saccharomyces cerevisiae.
    J Cell Biol. 1972 Jun;53(3):837-41 PMID: 4554992
  2. STUDIES ON THE YEAST NUCLEUS. I. THE ISOLATION OF NUCLEI.
    Biochim Biophys Acta. 1964 Sep 11;91:105-12 PMID: 14227256
  3. Letter: Fusion of two immunoglobulin-producing myeloma cells.
    Nature. 1973 Jul 6;244(5410):42-3 PMID: 4582485
  4. Purification and properties of flagellar outer doublet tubulin from Naegleria gruberi and a radioimmune assay for tubulin.
    J Biol Chem. 1974 Jun 10;249(11):3638-46 PMID: 4208665
  5. Duplication of spindle plaques and integration of the yeast cell cycle.
    Cold Spring Harb Symp Quant Biol. 1974;38:123-31 PMID: 4598635
  6. Ultrastructural localization of the high molecular weight proteins associated with in vitro-assembled brain microtubules.
    J Cell Biol. 1975 Apr;65(1):237-41 PMID: 1127013
  7. Immunofluorescence of mitotic spindles by using monospecific antibody against bovine brain tubulin.
    Science. 1975 Mar 14;187(4180):948-50 PMID: 1096300
  8. Membrane-bound tubulin in brain and thyroid tissue.
    J Biol Chem. 1975 Oct 10;250(19):7639-46 PMID: 1176440
  9. Cytoplasmic microtubules in tissue culture cells appear to grow from an organizing structure towards the plasma membrane.
    Proc Natl Acad Sci U S A. 1976 Mar;73(3):867-71 PMID: 1062799
  10. Griseofulvin interacts with microtubules both in vivo and in vitro.
    J Mol Biol. 1976 Apr 25;102(4):817-29 PMID: 781264
  11. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion.
    Eur J Immunol. 1976 Jul;6(7):511-9 PMID: 825377
  12. Immunochemical determination of tubulin.
    FEBS Lett. 1977 Jan 15;73(1):109-14 PMID: 838043
  13. Identity and polymerization-stimulatory activity of the nontubulin proteins associated with microtubules.
    Biochemistry. 1977 Jun 14;16(12):2598-605 PMID: 889779
  14. Radioimmunoassay for tubulin detection.
    FEBS Lett. 1977 Dec 1;84(1):97-100 PMID: 590533
  15. Immunocytochemical visualization of microtubules and tubulin at the light- and electron-microscopic level.
    J Cell Sci. 1977 Dec;28:283-301 PMID: 74378
  16. Combined immunofluorescence and high-voltage electron microscopy of cultured mammalian cells, using an antibody that binds to glutaraldehyde-treated tubulin.
    Proc Natl Acad Sci U S A. 1978 Jan;75(1):334-8 PMID: 415307
  17. Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells by electron microscopy and by immunofluorescence microscopy.
    Proc Natl Acad Sci U S A. 1978 Apr;75(4):1820-4 PMID: 417343
  18. Species-dependent immunological differences between vertebrate brain tubulins.
    Proc Natl Acad Sci U S A. 1978 Mar;75(3):1414-7 PMID: 77531
  19. Quantitation and characterization of antibody binding to tubulin.
    J Biol Chem. 1978 Sep 10;253(17):5980-4 PMID: 681333
  20. Myosins of secretory tissues.
    J Cell Biol. 1978 Jun;77(3):827-36 PMID: 150427
  21. Radioimmunoassay for tubulin: a quantitative comparison of the tubulin content of different established tissue culture cells and tissues.
    Cell. 1978 Aug;14(4):795-804 PMID: 688394
  22. Presence of NADPH-cytochrome P-450 reductase in rat liver Golgi membranes. Evidence obtained by immunoadsorption method.
    J Cell Biol. 1978 Nov;79(2 Pt 1):590-7 PMID: 214451
  23. Rat x rat hybrid myelomas and a monoclonal anti-Fd portion of mouse IgG.
    Nature. 1979 Jan 11;277(5692):131-3 PMID: 310519
  24. Microtubules in cultured cells; indirect immunofluorescent staining with tubulin antibody.
    Int Rev Cytol. 1980;63:59-95 PMID: 395133
  25. Differentiation antigens of the lymphocyte cell surface.
    Contemp Top Mol Immunol. 1977;6:83-116 PMID: 94279
  26. Three rat monoclonal antibodies to human C3.
    Immunology. 1980 Nov;41(3):503-15 PMID: 6161872
  27. Calcium lability of cytoplasmic microtubules and its modulation by microtubule-associated proteins.
    Proc Natl Acad Sci U S A. 1981 Feb;78(2):1037-41 PMID: 7015328
  28. Multiple alpha- and beta-tubulin genes in Chlamydomonas and regulation of tubulin mRNA levels after deflagellation.
    Cell. 1981 Apr;24(1):81-8 PMID: 6263492
  29. Purification of yeast tubulin by self-assembly in vitro.
    Biochemistry. 1981 Jun 9;20(12):3629-33 PMID: 7020758
  30. High resolution light and electron microscopic localization of tubulin with the IGS (immuno gold staining) method.
    Cell Biol Int Rep. 1981 Sep;5(9):889-99 PMID: 7028278
  31. Preparation of monoclonal antibodies: strategies and procedures.
    Methods Enzymol. 1981;73(Pt B):3-46 PMID: 7300683
  32. Expression of spleen cell immunoglobulin phenotype in hybrids with myeloma cell lines.
    Somatic Cell Genet. 1981 Nov;7(6):657-66 PMID: 7323946
  33. Transplantable immunoglobulin-secreting tumours in rats. Purification and chemical characterization of four kappa chains from LOU-Wsl rats.
    Eur J Biochem. 1972 Dec 4;31(2):354-9 PMID: 4630505
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.93.3.576
Published
1982-06-00
Pages
576-82
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2112140
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.93.3.576 PMC Full Text View journal details
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