Immunogen: Peptide in C-terminus of human CD33
Clonality: Monoclonal
Isotype: IgG2b
Host: Rat
Reactivity: Human

Overview

Monoclonal antibody 24F3 reacts with human CD33. In initial testing on lysates of HEK293 cells that overexpressed myc-tagged human CD33 isoforms, it preferentially recognized CD33m when used as a detection antibody on Western blots or a capture antibody for immunoprecipitation. The antibody was also shown to work in immunocytochemical applications, staining HEK cells transfected with human CD33-3 or CD33m.

Monoclonal antibody 24F3 recognizes human CD33m overexpressed in HEK cells. A) Western blots of lysates of HEK293 cells transfected with myc-tagged human CD33 isoforms or empty vector (mock). Bands representing all three human CD33 isoforms are seen in a blot probed with an anti-myc antibody. CD33-3 and CD33M are seen in a blot probed with Abcam anti-CD33 (134115), which detects an epitope within the first N-terminal 100 amino acids, missing in CD33m. By contrast, 24F3 preferentially detects CD33m. B) Antibody 24F3 also preferentially recognizes CD33m when used as a capture antibody for immunoprecipitation. Left blot, cell lysates immunoprecipitated with 24F3; right blot, cell lysates (input for immunoprecipitation). Blots probed with anti-myc antibody 9E10. *, non-specific bands. [Courtesy of Christian Haass' laboratory.]

Monoclonal antibody 24F3 recognizes human CD33 overexpressed in HEK cells. HEK cells transfected with human CD33 (CD33-3 or CD33m) or mouse (CD33m) isoforms were fixed, permeabilized, then incubated with 24F3 followed by a fluorescently conjugated secondary antibody. Cultures were counterstained with the nuclear stain DAPI (blue). Note 24F3 immunoreactivity (magenta) in the human CD33-transfected cells. As expected, this antibody did not recognize mouse CD33. [Courtesy of Christian Haass' laboratory.]

Generation and epitope mapping

This antibody was generated in rats immunized with a peptide in the C-terminal region of human CD33 that is present in all three isoforms, coupled to ovalbumin. Epitope mapping has not been reported.

Specificity

Based on the amino acid sequence of the peptide immunogen, clone 24F3 is expected to react with all three isoforms of the human protein. However, in initial testing, 24F3 preferentially recognized the CD33m isoform in western-blot and immunoprecipitation applications. The antibody is not expected to react with mouse CD33.

Validation

No data.

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References

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Further Reading

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