DAP12 Antibody [E15J7] | Primary Antibodies

Application: Reactivity: Rat, Mouse

Usage Information

Dilution
WB1:1000 IP1:30 IHC1:2000 IF1:50 FCM1:50

Application
WB, IP, IHC, IF, FCM

Reactivity
Rat, Mouse

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
12 kDa 12 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Datasheet & SDS

Biological Description

Specificity
DAP12 Antibody [E15J7] detects endogenous levels of total DAP12 protein.

Clone
E15J7

Synonym(s)
Dap12, Karap, Tyrobp, TYRO protein tyrosine kinase-binding protein, DNAX-activation protein 12, Killer-activating receptor-associated protein, KAR-associated protein

Background
DAP12 (TYROBP/KARAP) is a disulfide‑linked homodimeric transmembrane adaptor protein expressed in NK cells, myeloid cells, and other hematopoietic lineages, where it provides immunoreceptor tyrosine‑based activation motif (ITAM)–dependent signaling capacity to a broad set of ligand‑binding receptors. Each DAP12 chain contains a short extracellular region, a transmembrane segment with a negatively charged aspartate that pairs with basic residues in partner receptor transmembrane domains, and a cytoplasmic tail with a dual‑tyrosine ITAM that becomes phosphorylated by Src family kinases when associated receptors engage their ligands. Phosphorylated ITAM tyrosines recruit Syk (and ZAP‑70 in NK cells), which then autophosphorylates and phosphorylates scaffold proteins such as LAT and SLP‑76, leading to assembly of signaling complexes that include PI3K, PLCγ, Vav, Grb2–SOS, and Gads and propagate signals to AKT, ERK, PKC, calcium mobilization, and actin cytoskeleton rearrangement. DAP12 associates with multiple activating receptors, including TREM‑1 and TREM‑2 on myeloid cells, certain killer cell activating receptors on NK cells, MDL‑1, and SIRPβ1, so the same ITAM‑based module can couple diverse pattern‑recognition or immunoregulatory inputs to a conserved activation cascade. Ligation of TREM‑1–DAP12 on monocytes and neutrophils induces robust production of pro‑inflammatory cytokines and chemokines, generation of reactive oxygen species, degranulation, and phagocytosis, and TREM‑1–DAP12 blockade in experimental sepsis models reduces inflammatory mediator production and improves survival, highlighting this axis as an amplifier of innate inflammatory responses. TREM‑2–DAP12 signaling in macrophages, dendritic cells, osteoclasts, and microglia promotes phagocytosis of bacteria and cell debris, supports differentiation and function of osteoclasts, and can inhibit TLR‑driven inflammatory signaling, indicating context‑dependent roles that range from activation to negative feedback on inflammation. Mutations in DAP12 and TREM-2 are associated with distinct bone and neurological phenotypes, such as those seen in Nasu–Hakola disease, and influence microglial responses during neurodegeneration, as impaired DAP12-dependent signaling disrupts osteoclast-driven bone remodeling and alters microglial phagocytic and inflammatory activities.

Background

DAP12 (TYROBP/KARAP) is a disulfide‑linked homodimeric transmembrane adaptor protein expressed in NK cells, myeloid cells, and other hematopoietic lineages, where it provides immunoreceptor tyrosine‑based activation motif (ITAM)–dependent signaling capacity to a broad set of ligand‑binding receptors. Each DAP12 chain contains a short extracellular region, a transmembrane segment with a negatively charged aspartate that pairs with basic residues in partner receptor transmembrane domains, and a cytoplasmic tail with a dual‑tyrosine ITAM that becomes phosphorylated by Src family kinases when associated receptors engage their ligands. Phosphorylated ITAM tyrosines recruit Syk (and ZAP‑70 in NK cells), which then autophosphorylates and phosphorylates scaffold proteins such as LAT and SLP‑76, leading to assembly of signaling complexes that include PI3K, PLCγ, Vav, Grb2–SOS, and Gads and propagate signals to AKT, ERK, PKC, calcium mobilization, and actin cytoskeleton rearrangement. DAP12 associates with multiple activating receptors, including TREM‑1 and TREM‑2 on myeloid cells, certain killer cell activating receptors on NK cells, MDL‑1, and SIRPβ1, so the same ITAM‑based module can couple diverse pattern‑recognition or immunoregulatory inputs to a conserved activation cascade. Ligation of TREM‑1–DAP12 on monocytes and neutrophils induces robust production of pro‑inflammatory cytokines and chemokines, generation of reactive oxygen species, degranulation, and phagocytosis, and TREM‑1–DAP12 blockade in experimental sepsis models reduces inflammatory mediator production and improves survival, highlighting this axis as an amplifier of innate inflammatory responses. TREM‑2–DAP12 signaling in macrophages, dendritic cells, osteoclasts, and microglia promotes phagocytosis of bacteria and cell debris, supports differentiation and function of osteoclasts, and can inhibit TLR‑driven inflammatory signaling, indicating context‑dependent roles that range from activation to negative feedback on inflammation. Mutations in DAP12 and TREM-2 are associated with distinct bone and neurological phenotypes, such as those seen in Nasu–Hakola disease, and influence microglial responses during neurodegeneration, as impaired DAP12-dependent signaling disrupts osteoclast-driven bone remodeling and alters microglial phagocytic and inflammatory activities.

References
https://pubmed.ncbi.nlm.nih.gov/12570670/ https://pubmed.ncbi.nlm.nih.gov/12569153/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%