16-9039-82

antibody from Invitrogen Antibodies

Targeting: TLR3 CD283

Provider product page for 16-9039-82

Flow cytometry

Blocking/Neutralizing

Other assay

Antibody data

Antibody Data
Antigen structure
References [6]
Comments [0]
Validations
Flow cytometry [1]
Other assay [4]

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Product number: 16-9039-82 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD283 (TLR3) Monoclonal Antibody (TLR3.7), Functional Grade, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The TLR3.7 monoclonal antibody reacts with human Toll-like receptor 3 (TLR3). To date, at least twelve members of the Toll-like receptor family have been identified. This family of type I transmembrane proteins is characterized by an extracellular domain with leucine-rich repeats and a cytoplasmic domain with homology to the type I IL-1 receptor. In the innate immune response, TLRs recognize molecular patterns associated with microbial pathogens and induce antimicrobial activity. TLR3 recognizes double-stranded (ds)RNA, induces the activation of NF-kappaB through MyD88-dependent and -independent pathways, and the production of type I interferons (IFNs). TLR3.7 suppressed poly(I):poly(C)-mediated IFN-beta production by human fibroblasts naturally expressing TLR3 on their surface. Applications Reported:The TLR3.7 antibody has been reported for use in flow cytometric analysis. It has also been reported in in vitro inhibition of ligand binding. For optimal detection of TLR3 with this mAb by flow cytometry, PE-conjugated or Biotin-conjugated TLR3.7 (revealed with SAv-PE) are recommended. Applications Tested: This TLR3.7 antibody has been tested by intracellular staining and flow cytometric analysis of A549 and MRC5 cell lines to detect the very low level of this antigen on the surface. This can be used at less than or equal to 2 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. torage and handling: Use in a sterile environment. Filtration: 0.2 µm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level:Less than 0.001 ng/µg antibody, as determined by LAL assay. Aggregation:Less than 10%, as determined by HPLC.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: TLR3.7
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: 4° C

TLR3 protein structure - 16-9039-82 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 IRF1 regulates TRL2 and TLR3 signaling. (A) Parent and IRF1 KO cells were treated with poly I:C (1 mug/mL) for 6 h and IRF1 independent ISG expression was examined by RT-qPCR comparative C q method (2 -DeltaDelta Cq ). Data represent mean +- SEM from three independent experiments. (B) Parent and IRF1 KO cells were processed for confocal microscopy to examine localization of TLR3. Antibody against EEA1 was used to examine localization of early endosomes. Images from individual channels for TLR3 (green), EEA1 (red), and nuclei (blue) are shown. Right panels show images with merged channels with magnified view as inset. (C,D) Parent and IRF1 KO cells were treated with the respective TLR2 and TLR4 agonists, Pam2CSK4 at 1ng/ml (C) , or LPS at 10 ng/ml (D) and activation of NF-kB pathway was examined by western blot with anti-phospho-NFKBIA (S32). * Statistically significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Interleukin-1 receptor 1 is highly expressed on infra-patellar pad derived fibroblasts. Fibroblasts isolated from the infra-patellar fat pad and synovial membrane (both n = 5) were cultured in vitro and the relative gene expression of TLR1-10, RIG-1, RAGE and IL-1R1 quantified by qRT-PCR. ( a ) TLR expression was restricted mainly to TLR3 and TLR4 with little or no expression of other TLRs. ( b ) There was no detectable expression of RAGE but RIG-1 was expressed at levels comparable to TLR3 and TLR4. In contrast IL-1R1 expression was 100-150 fold greater than other receptors. Results are normalised to GAPDH as a loading control. Data is presented as mean +- standard error of the mean. Fibroblasts isolated from the infra-patellar fat pad were investigated for the expression of TLR3, TLR4 and IL-1R1 protein by Western Blotting (n = 4) ( c ) and flow cytometry (n = 6) ( d ). Cells expressed TLR3 and TLR4 at comparable levels at the cell surface. In contrast, expression of IL-1R1 was significantly higher than other receptors (red traces). beta-actin was used as a loading control (c) . Unstained cells (filled traces) and IgG controls (black traces) were used as flow cytometry controls (d) .