TLR9 Antibody - #DF2970

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Product: TLR9 Antibody
Catalog: DF2970
Description: Rabbit polyclonal antibody to TLR9
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Horse, Rabbit, Dog
Mol.Wt.: 117kD,140kD(full), 45kD,75kD(cleaved); 116kD(Calculated).
Uniprot: Q9NR96
RRID: AB_2840950

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 100ul $280 In stock
 200ul $350 In stock

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:1000-3000, IHC 1:500-1:2000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Horse(82%), Rabbit(82%), Dog(82%)
Clonality:
Polyclonal
Specificity:
TLR9 Antibody detects endogenous levels of total TLR9.
RRID:
AB_2840950
Cite Format: Affinity Biosciences Cat# DF2970, RRID:AB_2840950.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

CD 289; CD289; TLR 9; TLR9; TLR9_HUMAN; Toll like receptor 9; Toll like receptor 9 isoform A precursor; Toll like receptor 9 isoform B; Toll-like receptor 9;

Immunogens

Immunogen:
Uniprot:

Q9NR96(TLR9_HUMAN) >>Visit HPA database.

Gene(ID):
TLR9(54106)
Expression:
Q9NR96 TLR9_HUMAN:

Highly expressed in spleen, lymph node, tonsil and peripheral blood leukocytes, especially in plasmacytoid pre-dendritic cells. Levels are much lower in monocytes and CD11c+ immature dendritic cells. Also detected in lung and liver.

Sequence:
MGFCRSALHPLSLLVQAIMLAMTLALGTLPAFLPCELQPHGLVNCNWLFLKSVPHFSMAAPRGNVTSLSLSSNRIHHLHDSDFAHLPSLRHLNLKWNCPPVGLSPMHFPCHMTIEPSTFLAVPTLEELNLSYNNIMTVPALPKSLISLSLSHTNILMLDSASLAGLHALRFLFMDGNCYYKNPCRQALEVAPGALLGLGNLTHLSLKYNNLTVVPRNLPSSLEYLLLSYNRIVKLAPEDLANLTALRVLDVGGNCRRCDHAPNPCMECPRHFPQLHPDTFSHLSRLEGLVLKDSSLSWLNASWFRGLGNLRVLDLSENFLYKCITKTKAFQGLTQLRKLNLSFNYQKRVSFAHLSLAPSFGSLVALKELDMHGIFFRSLDETTLRPLARLPMLQTLRLQMNFINQAQLGIFRAFPGLRYVDLSDNRISGASELTATMGEADGGEKVWLQPGDLAPAPVDTPSSEDFRPNCSTLNFTLDLSRNNLVTVQPEMFAQLSHLQCLRLSHNCISQAVNGSQFLPLTGLQVLDLSHNKLDLYHEHSFTELPRLEALDLSYNSQPFGMQGVGHNFSFVAHLRTLRHLSLAHNNIHSQVSQQLCSTSLRALDFSGNALGHMWAEGDLYLHFFQGLSGLIWLDLSQNRLHTLLPQTLRNLPKSLQVLRLRDNYLAFFKWWSLHFLPKLEVLDLAGNQLKALTNGSLPAGTRLRRLDVSCNSISFVAPGFFSKAKELRELNLSANALKTVDHSWFGPLASALQILDVSANPLHCACGAAFMDFLLEVQAAVPGLPSRVKCGSPGQLQGLSIFAQDLRLCLDEALSWDCFALSLLAVALGLGVPMLHHLCGWDLWYCFHLCLAWLPWRGRQSGRDEDALPYDAFVVFDKTQSAVADWVYNELRGQLEECRGRWALRLCLEERDWLPGKTLFENLWASVYGSRKTLFVLAHTDRVSGLLRASFLLAQQRLLEDRKDVVVLVILSPDGRRSRYVRLRQRLCRQSVLLWPHQPSGQRSFWAQLGMALTRDNHHFYNRNFCQGPTAE

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Horse
82
Dog
82
Rabbit
82
Pig
76
Bovine
71
Sheep
71
Xenopus
59
Zebrafish
46
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9NR96 As Substrate

Site PTM Type Enzyme
S88 Phosphorylation
T202 Phosphorylation
Y345 Phosphorylation
Y419 Phosphorylation
S423 Phosphorylation
T642 Phosphorylation
T693 Phosphorylation
S696 Phosphorylation
K738 Acetylation
K789 Ubiquitination
Y888 Phosphorylation
S1004 Phosphorylation

Research Backgrounds

Function:

Key component of innate and adaptive immunity. TLRs (Toll-like receptors) control host immune response against pathogens through recognition of molecular patterns specific to microorganisms. TLR9 is a nucleotide-sensing TLR which is activated by unmethylated cytidine-phosphate-guanosine (CpG) dinucleotides. Acts via MYD88 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Controls lymphocyte response to Helicobacter infection (By similarity). Upon CpG stimulation, induces B-cell proliferation, activation, survival and antibody production.

PTMs:

Activated by proteolytic cleavage of the flexible loop between repeats LRR14 and LRR15 within the ectodomain. Cleavage requires UNC93B1. Proteolytically processed by first removing the majority of the ectodomain by either asparagine endopeptidase (AEP) or a cathepsin followed by a trimming event that is solely cathepsin mediated and required for optimal receptor signaling.

Subcellular Location:

Endoplasmic reticulum membrane>Single-pass type I membrane protein. Endosome. Lysosome. Cytoplasmic vesicle>Phagosome.
Note: Relocalizes from endoplasmic reticulum to endosome and lysosome upon stimulation with agonist. Exit from the ER requires UNC93B1. Endolysosomal localization is required for proteolytic cleavage and subsequent activation. Intracellular localization of the active receptor may prevent from responding to self nucleic acid.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Highly expressed in spleen, lymph node, tonsil and peripheral blood leukocytes, especially in plasmacytoid pre-dendritic cells. Levels are much lower in monocytes and CD11c+ immature dendritic cells. Also detected in lung and liver.

Subunit Structure:

Monomer and homodimer. Exists as a monomer in the absence of unmethylated cytidine-phosphate-guanosine (CpG) ligand. Proteolytic processing of an insertion loop (Z-loop) is required for homodimerization upon binding to the unmethylated CpG ligand leading to its activation (By similarity). Interacts with MYD88 via their respective TIR domains (By similarity). Interacts with BTK. Interacts (via transmembrane domain) with UNC93B1. Interacts with CD300LH; the interaction may promote full activation of TLR9-triggered innate responses (By similarity). Interacts with CNPY3 and HSP90B1; this interaction is required for proper folding in the endoplasmic reticulum. Interacts with SMPDL3B (By similarity).

Family&Domains:

Belongs to the Toll-like receptor family.

References

1). Mitochondrial DNA mediates immunoparalysis of dendritic cells in sepsis via STING signalling. CELL PROLIFERATION (PubMed: 36106559) [IF=8.5]

Application: WB    Species: Mice    Sample: BMDCs

FIGURE 4 Cytoplasmic mtDNA markedly activated STING signalling. BMDCs were treated with mtDNA (10 μg/ml), or transfected with vehicle or mtDNA (10 μg/ml) for 24 h, then were harvested for western blot and qPCR. (A) Representative blots and relative quantification protein levels of STING pathway (n = 3). (B) qPCR quantitation of IFN‐β (n = 7). (C) Representative images of nuclear translocation of IRF3 in BMDC treated as indicated. Scale bar: 5 μm. (D) Representative blots and relative quantification protein levels of TLR9 pathway (n = 3). (E) qPCR quantitation of TNF‐α, IL‐6 and VCAM‐1 (n = 6–7). (F) Representative blots and relative quantification protein levels of NLRP3 pathway (n = 3). (G) qPCR quantitation of IL‐1β, IL‐18 (n = 7). Data are shown as the mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001. BMDC, bone marrow derived dendritic cell; IFN, interferon; mtDNA, mitochondrial DNA; mtDNA (T), mtDNA transfection; Veh (T), vehicle transfection.
2). Inhibition of murine herpesvirus-68 replication by IFN-gamma in macrophages is counteracted by the induction of SOCS1 expression. PLOS Pathogens (PubMed: 30075008) [IF=6.7]

Application: WB    Species: mouse    Sample: macrophages

Fig 6. |TLR3 mediates the MHV-68-induced SOCS1 production. (A) BMMs were transfected with siRNAs against Tlr2, Tlr3, Tlr4, Tlr7, Tlr9 (si-TLR) or an irrelevant target (si-Control), respectively. Forty-eight hours post transfection, cells were infected with MHV-68 at MOI = 10. At 8 hpi, RNA and protein were then extracted. RT-qPCR was performed to determine SOCS1 mRNA levels and western blotting was performed to determine the protein expression levels of SOCS1 and the individual TLRs, respectively. The SOCS1 mRNA levels are expressed as values relative to the MHV-68 infected, si-Control transfected cells.
3). 25-HC promotes hepatocellular carcinoma metastasis through up-regulation of TLR4 dependent FABP4. American Journal of Cancer Research (PubMed: 31720079) [IF=5.3]

Application: WB    Species: Human    Sample: HepG2 cells

Figure 5 The migration of HepG2 cells induced by 25-HC is partly mediated by the up-regulation of TLR4. (A) HepG2 cells were treated with the indicated concentrations of 25-HC for 24 hours before mRNA was extracted and expressions of TLR2, TLR4, TLR5, TLR7 and TLR9 were examined by RT-qPCR. (B) HepG2 cells were treated with the indicated concentrations of 25-HC for 24 hours or treated with 25-HC at 10 μM for the indicated times before protein was collected to detect the TLR2, TLR4, TLR5, TLR7 and TLR9 expressions by Western blotting. HepG2 cells were transfected with siNC or siTLR4 or siTLR9 for 24 hours, (C) then cells were treated with the indicated concentrations of 25-HC for 24 hours before proteins were collected and the expressions of MMP1, MMP2, MMP3 and MMP9 were determined by Western blotting. (D) Or migratory ability of HepG2 cells after treated with 25-HC for 36 hours was determined by Transwell assay. Results were obtained from 3 independent experiments and are expressed as the means ± SEM. Statistical significance was determined by Student’s t-test. *P<0.05, ***P<0.001.
4). 25-HC decreases the sensitivity of human gastric cancer cells to 5-fluorouracil and promotes cells invasion via the TLR2/NF-κB signaling pathway. International Journal of Oncology (PubMed: 30664194) [IF=5.2]
5). Mitochondrial transcription factor B2 overexpression increases M2 macrophage infiltration via cytosolic mitochondrial DNA-stimulated Interleukin-6 secretion in ovarian cancer. Bioengineered (PubMed: 35577351) [IF=4.9]

Application: IF/ICC    Species: Human    Sample: OVISE and CAOV4 cells

Figure 3. Cytosolic mtDNA stress enhances the secretion of IL-6 by activating the nuclear factor-κB (NF-κB) signaling pathway A-B. qRT-PCR analysis for the mRNA expression of genes encoding TAM recruitment associated cytokines and chemokines including CCL2, CSF1, TGF-β, IL-6, and CCL22 in OVISE (a) and CAOV4 (b) cells 48 h after lentiviral mediated TFB2M overexpression. C-D. qRT-PCR (c) analysis of IL-6 transcription in cells and ELISA (d) assay of IL-6 secretion in the supernatants of cultured OVISE and CAOV4 cells with TFB2M overexpression alone, treated with DNase I alone or with combined TFB2M overexpression and DNase I treatment. E. qPCR was used to determine the copy number of mtDNA in the cytoplasm of OVISE and CAOV4 cells with TFB2M overexpression alone, treated with DNase I alone or with combined TFB2M overexpression and DNase I treatment. F. Immunofluorescent staining of TLR9 in OVISE and CAOV4 cells with or without TFB2M overexpression. The interactions between TLR9 and cytosolic mtDNA were indicated by white arrows. G-J. Western blot analyses were performed to detect the protein levels of TFB2M in whole cells and phospho-NF-κB p65 (p-p65) (Ser536) and p65 in the cytoplasm and nucleus of OVISE and CAOV4 cells with TFB2M overexpression alone or with combined ODN INH-18 (a TLR9 antagonist), DNase I or PDTC (an NF-κB inhibitor) treatment. Total TFB2M, nucleus p-p65, and p65 in each group were quantified using ImageJ software (h-j). Data shown are the mean ± SD. from three independent experiments. EV: empty vector. **p < 0.01, ***p < 0.001, n.s., not significant.
6). Electroacupuncture preconditioning protects against myocardial ischemia-reperfusion injury by modulating dynamic inflammatory response. Heliyon (PubMed: 37809701) [IF=4.0]

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