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AdipoGen Life Sciences
Flagellin (NLRC4 Mutant) (rec.)
215
CHF
CHF 215.00
In stock
AG-40B-0126-C01010 µgCHF 215.00
AG-40B-0126-30103 x 10 µgCHF 480.00
Figure 1: Flagellin (rec.) (AG-40B-0125) and Flagellin (NLRC4 Mutant) (rec.) (AG-40B-0126) induce CCL20-luciferase promoter through TLR5 in vitro.
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: 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.
Figure 2: Flagellin (rec.) (AG-40B-0125) and Flagellin (NLRC4 Mutant) (rec.) (AG-40B-0126) activate TLR5 in vivo.
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.
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.
Figure 3: Flagellin (rec.) (AG-40B-0125) activates NLRC4 inflammasome in bone-marrow mouse macrophages.
Method: Bone-marrow mouse macrophages (BMDMs) from WT or NLRC4 -/- mice are either left untreated, or incubated with streptolysin (SLO), or SLO with Flagellin (rec.) (AG-40B-0125) (=WT), or SLO with Flagellin (rec.) (TLR4 Mutant) (AG-40B-0126) at the indicated concentrations for 10min. Macrophages are then washed and incubated further for 2h before the cell extracts are prepared and immunoblotted for caspase-1 activation. p10 or p20 fragments are detected using the anti-Caspase-1 (p10) (mouse), mAb (Casper-2) (AG-20B-0044) or the anti-Caspase-1 (p20) (mouse), mAb (Casper-1) (AG-20B-0042), respectively.
Method: Bone-marrow mouse macrophages (BMDMs) from WT or NLRC4 -/- mice are either left untreated, or incubated with streptolysin (SLO), or SLO with Flagellin (rec.) (AG-40B-0125) (=WT), or SLO with Flagellin (rec.) (TLR4 Mutant) (AG-40B-0126) at the indicated concentrations for 10min. Macrophages are then washed and incubated further for 2h before the cell extracts are prepared and immunoblotted for caspase-1 activation. p10 or p20 fragments are detected using the anti-Caspase-1 (p10) (mouse), mAb (Casper-2) (AG-20B-0044) or the anti-Caspase-1 (p20) (mouse), mAb (Casper-1) (AG-20B-0042), respectively.
| Product Details | |
|---|---|
| Synonyms | FliC (NLRC4 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 small deletion at the C-terminus of the protein required for NLRC4 activation and is fused at the N-terminus to a His-tag. |
| Crossreactivity |
Human Mouse |
| Specificity |
Detected by human and mouse TLR5 (Toll-like receptor 5). Not detected by mouse NLRC4 (Nod-like receptor [NLR] family caspase activation and recruitment domain [CARD] domain-containing protein 4). |
| Biological Activity |
Activation of TLR5 in human epithelial cell assays based on NF-κB luciferase fusions, in vivo on expression of CCL20 or IL-6. TLR5 can be stimulated in vitro by Flagellin (NLRC4 mutant) (rec.) at a concentration range of 5-50ng/ml and in vivo at a starting concentration of 0.1-1µg/mouse. Does not activate NLRC4 as tested in bone-marrow mouse macrophages (in the presence of streptolysin O (SLO)). It can be used as negative control for NLRC4 signaling. |
| 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 |
Description
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 (NLRC4 Mutant)(rec.) is only detected by TLR5 not by NLRC4.
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