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

Overview

In initial testing, monoclonal antibody 14C2 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 recognized human DAP12 overexpressed in HEK293 cells in immunocytochemical (see below) and immunoprecipitation (not shown) applications.

Monoclonal antibody 14C2 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 14C2 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 14C2 or anti-myc (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.]

Monoclonal antibody 14C2 recognizes human DAP12 overexpressed in HEK293 cells. HEK293 cells transfected with TYROBP and untransfected controls were stained with 14C2 followed by an Alexa-647-conjugated secondary antibody, then counterstained with the nuclear marker DAPI. DAP12 immunoreactivity is shown in magenta, DAPI nuclear staining in blue. [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 14C2 recognizes mouse and human DAP12.

Validation

14C2 has been validated for western blotting and immunoprecipitation applications, using BV2 cells with  the Tyrobp gene knocked out.

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