Toll-like Receptor 9 Antibody [K21F2]

Application: Reactivity: Human, Mouse, Rat, Canine, Equine, Primate, Monkey

Usage Information

Dilution
WB1:400-1:1000 IP1:10-1:500 IHC1:400 IF1:10-1:500 FCM1:10 - 1:1000

Application
WB, IP, IHC, IF, FCM, ELISA

Reactivity
Human, Mouse, Rat, Canine, Equine, Primate, Monkey

Source
Mouse 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
115 kDa ~120 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
Toll-like Receptor 9 Antibody [K21F2] detects endogenous levels of total Toll-like Receptor 9 protein.

Clone
K21F2

Synonym(s)
CD289 antigen, CD289, TLR9, toll-like receptor 9

Background
TLR9 belongs to the Toll-like receptor family of pattern recognition receptors that orchestrate innate immune responses to microbial nucleic acids. Expressed primarily in plasmacytoid dendritic cells, B cells, and macrophages, TLR9 resides in the endoplasmic reticulum until ligand engagement triggers endosomal trafficking where cathepsin-mediated cleavage generates an active N-terminal ligand-binding fragment while retaining the C-terminal TIR signaling domain. Unmethylated CpG DNA motifs bind within the LRR horseshoe structure, inducing receptor dimerization and high-affinity recruitment of the MyD88 adaptor protein; this assembles the myddosome complex with IRAK4 kinase phosphorylating IRAK1/2, which then activates TRAF6 auto-ubiquitination. TRAF6 recruits the TAK1 kinase complex that splits signaling into two arms—activation of IKK leading to NF-κB nuclear translocation and transcription of TNF-α, IL-6, and IL-12, alongside MAPK cascades stabilizing AP-1 for enhanced cytokine production. In plasmacytoid DCs, a parallel pathway incorporates TRAF3 with IKKα to phosphorylate IRF7, driving robust type I interferon responses. UNC93B1 chaperones TLR9 trafficking, while AP-3 complex directs interferon-competent signaling to lysosome-related organelles distinct from early endosomal NF-κB compartments. TLR9 coordinates antiviral states through interferon-stimulated genes, matures antigen-presenting cells for adaptive immunity, and enhances phagocytosis via ROS production. Self-DNA immune complexes activate TLR9 in systemic lupus erythematosus, fueling autoreactive B cell expansion and interferon-driven pathology.

Background

TLR9 belongs to the Toll-like receptor family of pattern recognition receptors that orchestrate innate immune responses to microbial nucleic acids. Expressed primarily in plasmacytoid dendritic cells, B cells, and macrophages, TLR9 resides in the endoplasmic reticulum until ligand engagement triggers endosomal trafficking where cathepsin-mediated cleavage generates an active N-terminal ligand-binding fragment while retaining the C-terminal TIR signaling domain. Unmethylated CpG DNA motifs bind within the LRR horseshoe structure, inducing receptor dimerization and high-affinity recruitment of the MyD88 adaptor protein; this assembles the myddosome complex with IRAK4 kinase phosphorylating IRAK1/2, which then activates TRAF6 auto-ubiquitination. TRAF6 recruits the TAK1 kinase complex that splits signaling into two arms—activation of IKK leading to NF-κB nuclear translocation and transcription of TNF-α, IL-6, and IL-12, alongside MAPK cascades stabilizing AP-1 for enhanced cytokine production. In plasmacytoid DCs, a parallel pathway incorporates TRAF3 with IKKα to phosphorylate IRF7, driving robust type I interferon responses. UNC93B1 chaperones TLR9 trafficking, while AP-3 complex directs interferon-competent signaling to lysosome-related organelles distinct from early endosomal NF-κB compartments. TLR9 coordinates antiviral states through interferon-stimulated genes, matures antigen-presenting cells for adaptive immunity, and enhances phagocytosis via ROS production. Self-DNA immune complexes activate TLR9 in systemic lupus erythematosus, fueling autoreactive B cell expansion and interferon-driven pathology.

References
https://pubmed.ncbi.nlm.nih.gov/25309543/

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

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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%