Flagellin, TLR5 Ligand Summary
| Immunogen |
Flagellin FliC from Salmonella Tyhimurium (494 amino acid protein) is a highly conserved molecule among both gram-negative and gram-positive bacteria. |
Applications/Dilutions
| Dilutions |
- Functional
- In vitro assay
- Ligand Activation
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| Application Notes |
Activation of TLR5. Flagellin is a potent stimulator of innate immune responses in a number of eukaryotic cells and organisms, including both mammals and plants. In mammals, flagellin is recognized by TLR5 and triggers defense responses both systemically and at epithelial surfaces. Flagellin induces the activation of NF-KB and the production of cytokines and nitricoxide depending on the nature of the TLR5 signaling complex. Stimulation of TLR5 has been reported with 10-100 ng/ml. Use in functional reported in scientific literature (PMID: 24154872) Use in In vitro assay reported in scientific literature (PMID 25466255). Use in Ligand activation reported in scientific literature (PMID 25957979) |
| Publications |
Read Publications using
NBP2-25289 in the following applications:
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Reactivity Notes
Human reactivity reported in scientific literature (PMID: 24154872). Mouse reactivity reported in scientific literature (PMID: 25466255)
Packaging, Storage & Formulations
| Storage |
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer |
Prepare dilutions by adding the appropriate amount of PBS or tissue culture media. Contents: 10 ug in 100 ul of PBS with 10% glycerol |
| Concentration |
Please see the protocols for proper use of this product. If no protocol is available, contact technical services for assistance. |
Background
Flagellin ligand activity on TLR5/NF-kB SEAPorter™ HEK 293 cell line (Cat. No. NBP2-26277 [TLR5 positive; TLR2/TLR4 negative]) and NF-kB SEAPorter™ RAW 264.7 cell line (Cat. No. NBP2-26261 [TLR5 negative; TLR2/TLR4 positive]). NBP2-26277 is a stably co-transfected cell line that expresses full-length human Toll-like receptor 5 (TLR5) and the secreted alkaline phosphatase (SEAP) reporter gene under the transcriptional control of an NF-kB response element. NBP2-26261 is a stably transfected cell line which expresses the NF-kB/SEAP reporter gene. NBP2-26277 and NBP2-26261 cells were plated in 96-well plates at 5 x 10^4 cells/well and 8.5 x 10^4 cells/well, respectively. After 16 h, cells were stimulated with various amounts of Flagellin (NBP2-25289) as well as 500 ng/ml LPS (NBP2-26216) and 50 ng/ml Pam3CSK4 for 24 h. SEAP was analyzed using the SEAPorter Assay Kit (Cat. No. NBP2-25285). Please see figures 1 and 2.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Support products are
guaranteed for 6 months from date of receipt.
Product General Protocols
View specific protocols for Flagellin, TLR5 Ligand (NBP2-25289):
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
FAQs for Flagellin, TLR5 Ligand (NBP2-25289). (Showing 1 - 2 of 2 FAQs).
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Can this ligand bind not only to Human TLR5 but also to Mouse TLR5. Also, which other species can the ligand bind to?
- We test the Flagellin on human, we have not evaluated the protein on other species. The Flagellin was purified from was purified from S. typhimurium (TH4778 (fljB-fliC+) as described in the Hayashi reference (2001). Mammalian and other species have immune responses to Flagellin, for example responses have been described in fish and plants, please see this link. Please note that this is general scientific literature, rather than literature citing the NBP2-25289 product. However, based on the scientific literature, it is likely that the recombinant flagellin would bind to other species besides human. However, the researcher would need to empirically determine if the NBP2-25289 recombinant protein was suitable for their non-human model.
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What kind of cells is this recombinant product made in?
- The recombinant protein was expressed in E coli.
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