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April 1, 1986Journal of Histochemistry & CytochemistryOpen Access

Double-enzyme conjugates, producing an intermediate color, for simultaneous and direct detection of three different intracellular immunoglobulin determinants with only two enzymes.

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Authors

ECEric ClaassenVrije Universiteit Amsterdam DBD. M. BoorsmaGTx (United States)NKN. KorsAmsterdam University Medical Centers

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Implication

Experimental study demonstrates direct detection of three intracellular immunoglobulins in spleen sections, indicating a streamlined approach for multi-target cytochemical staining.

Key Points

To develop a rapid, direct cytochemical method for detecting three different intracellular immunoglobulin determinants simultaneously in a single tissue section using only two enzymes.
Synthesized a double-enzyme conjugate by covalently coupling alkaline phosphatase (AP) to horseradish peroxidase (HRP).
Coupled three distinct antigens or antibodies separately to AP, HRP, and the hybrid AP-HRP conjugate.
Treated spleen sections from immunized animals with a mixture of all three conjugates in a one-step incubation followed by a sequential two-step AP and HRP cytochemical development.
Cytochemical reaction of the AP-HRP conjugate produced a stable intermediate violet product distinguishable from single AP (blue) and HRP (red) staining.
The triple staining method successfully resolved three distinct antibody-forming cell populations and three intracellular antibody classes simultaneously as red, blue, and violet cells in single sections.

Cite This Study

Claassen et al. (1986) studied this question.

synapsesocial.com/papers/6a9aefe56210ce96fed15a31 https://doi.org/10.1177/34.4.2419394

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Implication

Experimental study demonstrates direct detection of three intracellular immunoglobulins in spleen sections, indicating a streamlined approach for multi-target cytochemical staining.

Double-enzyme conjugates, producing an intermediate color, for simultaneous and direct detection of three different intracellular immunoglobulin determinants with only two enzymes.

Discussion

Member takes

Implication

Key Points

Cite This Study

Double-enzyme conjugates, producing an intermediate color, for simultaneous and direct detection of three different intracellular immunoglobulin determinants with only two enzymes.

Discussion

Member takes

Implication

Key Points

Cite This Study

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