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AdipoGen Life Sciences
Flagellin (TLR5 Mutant) (rec.)
Method: Caco-2 cells transfected with plasmid encoding CCL20-LUC plasmid are activated for 24h with the indicated concentrations of Flagellin (rec.) (AG-40B-0125) and (NLRC4 Mutant) (rec.) (AG-40B-0126). As a negative control, Flagellin (TLR5 Mutant) (rec.) (AG-40B-0127) is used. Luciferase assays are performed on cell lysates and luminescence is normalized to mock-treated cells.
Method: Mice WT are injected i.v. with 1µg of Flagellin (rec.) (AG-40B-0125)(=WT) or Flagellin (NLRC4 Mutant) (rec.) (AG-40B-0126). After 2 hours, levels of CCL20 in the serum are measured by ELISA. As control, WT mice are injected i.v. with 1µg of Flagellin (TLR5 Mutant) (rec.) (AG-40B-0127). PBS injected mice are used as negative control.
Product Details | |
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Synonyms | FliC (TLR5 Mutant) |
Product Type | Protein |
Properties | |
Source/Host | E. coli |
Sequence |
The native flagellin [FliC] from Salmonella enterica Serovar Typhimurium strain ATCC14028 (aa 1-495) contains a substitution of amino acids 89-96 required for TLR5 signaling by a motif from a non-signaling flagellin and is fused at the N-terminus to a His-tag. |
Crossreactivity |
Human Mouse |
Specificity |
Not detected by human and mouse TLR5 (Toll-like receptor 5). |
Biological Activity |
Does not activate TLR5 in human epithelial cell assays based on NF-κB luciferase fusions, in vivo on expression of CCL20 or IL-6. It can be used as negative control for TLR5 signaling assays. |
Purity | ≥95% (SDS-PAGE) |
Endotoxin Content | <0.01EU/μg purified protein (LAL test). |
Concentration | 0.1mg/ml after reconstitution. |
Reconstitution | Reconstitute with 100μl sterile water. |
Formulation | Lyophilized. Contains PBS. |
Other Product Data |
NCBI link NP_460912.1: Flagellin [Salmonella enterica subsp. enterica serovar Typhimurium str. LT2] |
Shipping and Handling | |
Shipping | BLUE ICE |
Short Term Storage | +4°C |
Long Term Storage | -20°C |
Handling Advice |
After reconstitution, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles. Centrifuge lyophilized vial before opening and reconstitution. |
Use/Stability | Stable for at least 6 months after receipt when stored at -20°C. |
Documents | |
MSDS | Download PDF |
Product Specification Sheet | |
Datasheet | Download PDF |
Flagellin is the subunit protein which polymerizes to form the filaments of bacterial flagella. It activates the innate immune system through the receptor Toll-like Receptor 5 (TLR5) or the intracellular NLRC4 protein. The Flagellin (TLR5 Mutant) (rec.) cannot be detected by human and mouse TLR5. It has not been tested on NLRC4.
- Deletion of Flagellin’s Hypervariable Region Abrogates Antibody-Mediated Neutralization and Systemic Activation of TLR5-Dependent Immunity: C. Nempont, et al.; J. Immunol. 181, 2036 (2008)
- Mucosal Administration of Flagellin Protects Mice from Streptococcus pneumoniae Lung Infection: N. Munoz, et al.; Infect. Immun. 78, 4226 (2010)
- A Toll-Like Receptor 5 Agonist Improves the Efficacy of Antibiotics in Treatment of Primary and Influenza Virus-Associated Pneumococcal Mouse Infections: R. Porte, et al.; Antimicrob. Agents Chemother. 59, 6064 (2015)
- Recombinant flagellins with deletions in domains D1, D2, and D3: Characterization as novel immunoadjuvants: M.E. Biedma, et al.; Vaccine 37, 652 (2019)