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

Overview

In initial testing, monoclonal antibody 1C8 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.

Monoclonal antibody 1C8 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 1C8 hybridoma supernatant. A band (arrow) detected in the wild-type cells is absent from the knockout cells. B) Immunoprecipitation using 1C8 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.]

Antibody 1C8 recognizes human DAP12 expressed in HEK293 cells. HEK293 cells were stably transfected with human DAP12 and TREM2, a microglial receptor that activates DAP12. Immunoprecipitation of cell lysates using 1C8 as the capture antibody; blot probed with rabbit monoclonal anti-DAP12 (Cell Signaling #12492). HEK293 cells transfected with empty vector (mock) served as a negative control. 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 1C8 recognizes mouse and human DAP12.

Validation

1C8 has been validated for western blotting and immunoprecipitation applications, using BV2 cells with  the Tyrobp gene knocked out.

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