Immunogen: Peptide in C-terminus of human DAP12
Clonality: Monoclonal
Isotype: IgG2c
Host: Rat
Reactivity: Human; Mouse

Overview

In initial testing, monoclonal antibody 4E12 recognized DAP12 endogenously expressed in the BV2 mouse microglial cell line, when used as a detection antibody for western blotting or a capture antibody for immunoprecipitation. The antibody also immunoprecipitated human DAP12 overexpressed in HEK293 cells (not shown).

Monoclonal antibody 4E12 recognizes mouse DAP12 endogenously expressed in BV2 cells. A) Western blot of membrane extracts from wild-type cells (BV2) or cells in which the Tyrobp gene was knocked out (BV2 DAP12 KO), probed with 4E12 hybridoma supernatant. The DAP12 band (arrow) detected in the wild-type cells is absent from the knockout cells. The intervening lane was masked by the Alzforum curator. B) Immunoprecipitation using 4E12 or anti-Flag (negative control) as capture antibodies; blot probed with rabbit monoclonal anti-DAP12 (Cell Signaling #12492). The arrow indicates DAP12; asterisks show non-specific bands. [Courtesy of Christian Haass' laboratory.]

Generation and epitope mapping

This antibody was generated in rats immunized with a peptide in the C-terminus of human DAP12, coupled to ovalbumin. Epitope mapping has not been reported.

Specificity

Antibody 4E12 recognizes mouse and human DAP12.

Validation

4E12 has been validated for western blotting and immunoprecipitation applications, using BV2 cells with  the Tyrobp gene knocked out.

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