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Updated: May 31, 2026

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Published on: October 25, 2018

[Human CD96 gene cloning, expression and identification].

Jian-ming Zeng¹, Fei Liu, Ping-hai Tan

¹Department of Laboratory Medicine, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou 510120, China.

Nan Fang Yi Ke Da Xue Xue Bao = Journal of Southern Medical University

|July 19, 2011

Summary

This summary is machine-generated.

Researchers successfully cloned and expressed the human CD96 outer membrane domain (hCD96om) in E. coli. A specific rabbit polyclonal antibody against hCD96om was also developed for potential diagnostic applications in acute myeloid leukemia.

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Area of Science:

Molecular Biology
Immunology
Biotechnology

Background:

CD96 is a cell surface receptor implicated in immune responses and cancer progression.
Understanding CD96 function requires specific reagents for its detection and study.
Acute myeloid leukemia (AML) is a hematological malignancy where CD96 may play a role.

Purpose of the Study:

To construct and express the human CD96 outer membrane domain (hCD96om) in prokaryotic cells.
To generate a specific rabbit polyclonal antibody against hCD96om.

Main Methods:

hCD96om gene was amplified from AML patient peripheral blood via RT-PCR.
The gene was inserted into the pET32a(+) prokaryotic expression vector.
Recombinant protein expression in E. coli BL21(DE3) was induced and identified by Western blotting.
Rabbit polyclonal antibody production involved immunization with the purified fusion protein.

Main Results:

hCD96om protein was successfully expressed in E. coli BL21(DE3) as inclusion bodies (approx. 37 kD).
Western blotting confirmed the antibody's specific reaction with the expressed hCD96om fusion protein.
The generated antiserum also recognized a ~70 kD protein in human leukemia cell line HL-60.

Conclusions:

The human CD96 gene's outer membrane domain was successfully cloned and expressed in E. coli.
A specific rabbit polyclonal antibody against hCD96om was successfully prepared.