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. 1995 Oct;104(4):309-16.
doi: 10.1007/BF01464327.

Monoclonal antibody against the glutaraldehyde-conjugated polyamine, spermine

K Fujiwara  1 , Y Masuyama
Affiliations

Monoclonal antibody against the glutaraldehyde-conjugated polyamine, spermine

K Fujiwara et al. Histochem Cell Biol. 1995 Oct.

Abstract

We developed a mouse monoclonal antibody (ASPM-29, mAb) against spermine (Spm) conjugated to human serum albumin (HSA) using glutaraldehyde-sodium borohydride, for applications in immunocytochemistry (ICC). The antibody specificity was evaluated by an enzyme-linked immunosorbent assay (ELISA) binding test, simulating the ICC of tissue sections. ASPM-29 showed an almost equal immunoreactivity to Spm and spermidine (Spd) but no reactivity to any of the other polyamine (PA)-related compounds tested. By use of this antibody, indirect immunoperoxidase staining was observed in different tissues fixed with glutaraldehyde in combination with borohydride reduction. In contrast, immunoreactivity was quite low in tissues fixed only with glutaraldehyde. Absorption controls indicated that the immunostaining could be completely inhibited by 50 micrograms/ml of Spm or Spd and partially inhibited by N-acetylspermine (Ac-Spm), N1-acetylspermidine (N1-Ac-Spd), or N8-acetylspermidine (N8-Ac-Spd), but was hardly inhibited at all by other PA-related compounds or amino acids. The reactivity of the antibody with Spm conjugated on wells in an ELISA plate was inhibited by micromolar concentrations of Spm, Spd, Ac-Spm, N1-Ac-Spd, or N8-Ac-Spd, in decreasing order, but not by other small molecules. Dense ICC staining was observed in the paranuclear and basal cytoplasm of acinar cells of rat pancreas, submandibular gland and paratid gland, these results being in complete agreement with our recent ICC methods using other mAbs produced against N-(gamma-male-imidobutyryloxy) succinimide-conjugated Spm.

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References

    1. Biochem Biophys Res Commun. 1975 Dec 1;67(3):1070-7 - PubMed
    1. J Immunol Methods. 1983 Jul 15;61(2):217-26 - PubMed
    1. J Histochem Cytochem. 1966 Apr;14(4):291-302 - PubMed
    1. Histochem J. 1987 Dec;19(12):643-50 - PubMed
    1. Brain Res. 1984 Feb 27;294(1):161-5 - PubMed