Human IL-17/IL-17A Luciferase - (LUCPorter™) Stable Reporter Cell Line

Product Discontinued

Human IL-17/IL-17A Luciferase - (LUCPorter™) Stable Reporter Cell Line Summary

• Description: The IL-17A Prom/LUCPorter™ HEK 293 cell line can be used for screening of agonists, antagonists, or signaling inhibitors related to IL-17A induction. Note: The IL-17A Prom/LUCPorter™ HEK 293 cell line is developed as a semi-constitutively active reporter cell line that can be utilized for screening either agonists or antagonists for IL-17A induction. Induction of the IL-17A promoter by RORgt has been demonstrated and also shown in our test (figure 1) . In the test, RORg or RORgt significantly mediated activation of the IL-17A promoter which was quantified by luciferase activity (figure 1). Furthermore, Repression of the IL-17A promoter by a novel isoform of RORgt (RORgt[del. 5-8]), which lacks the hinge-encoding exons 5-8, has been demonstrated and also shown in our test (Figure 2).
  
  Contents: 3~4 x 10^6 cells
  Biosafety Level: 2
• Immunogen: IL-17A Prom/LUCPorter™ HEK 293 is a stably transfected cell line which expresses an optimized Renilla luciferase reporter gene (RenSP) under the transcriptional control of the IL-17A promoter.
• Target Species: Human
• Specificity: IL-17A Prom/LUCPorter ™
• Selection Agent: Puromycin.
• Growth Properties: Adherent Morphology: Epithelial
• RCL Type: Luciferase - (LUCPorter™)
• Host: HEK293
• Gene: IL17A

Applications/Dilutions

• Dilutions:
  • Ligand Activation
• Agonist:
  • ROR gamma Plasmid NBP2-25278
  • ROR gamma(t) Plasmid NBP2-25279
  • ROR gamma (t) [del. 5-8] Plasmid NBP2-25280
• Antagonist:
  • Thymoquinone, IL-17A Inhibitor (IL17 Inhibitor) NBP2-26241
  • Plumbagin (IL17 Inhibitor) NBP2-26242
  • Celastrol, IL-17A (IL-17A Inhibitor) NBP2-29362
• Readout System:
  • LightSwitch Luciferase Assay Reagent NBP2-25287
  • LightSwitch Luciferase Assay Reagent NBP2-32066

Packaging, Storage & Formulations

• Storage: Store in gas phase of liquid nitrogen.
• Reconstitution Instructions: Complete Growth Medium: DMEM with 4.5 g/L glucose + 10% FBS + 4 mM L-glutamine + 1 mM sodium pyruvate + 100 units/ml penicillin + 0.1 mg/ml streptomycin + 3 ug/ml puromycin.

Details for Array

• Type: Cell

Notes

Assume all cultures are hazardous since they may harbor latent viruses or other organisms that are uncharacterized. The following safety precautions should be observed.
- Use pipette aids to prevent ingestion and keep aerosols down to a minimum.
- No eating, drinking or smoking while handling the IL-17A reporter cell line.
- Wash hands after handling the IL-17A reporter cell line and before leaving the lab.
- Decontaminate work surface with disinfectant or 70% ethanol before and after working with the cell line.
- All waste should be considered hazardous.
- Dispose of all liquid waste after each experiment and treat with bleach.

Alternate Names for Human IL-17/IL-17A Luciferase - (LUCPorter™) Stable Reporter Cell Line

• CTLA8
• CTLA-8
• CTLA8cytotoxic T-lymphocyte-associated serine esterase 8
• Cytotoxic T-lymphocyte-associated antigen 8
• IL17
• IL-17
• IL17A
• IL-17A
• IL-17Acytotoxic T-lymphocyte-associated protein 8
• IL-17CTLA-8
• IL17interleukin-17A
• interleukin 17 (cytotoxic T-lymphocyte-associated serine esterase 8)
• interleukin 17A

Background

Interleukin 17 (IL-17) is a pro-inflammatory cytokine produced by TCRalpha/beta+ CD4 CD8 thymocytes, as well as by activated CD4+ and CD4+ CD45RO+ memory T cells (1). IL-17 stimulates the expression of TNFalpha, IL-1 beta, IL-6, and IL-8 (2). In addition, IL-17 is involved in the induction of inducible nitric oxide synthase and cyclooxygenase 2 in chondrocytes (3). High levels of IL-17 are associated with several chronic inflammatory diseases including rheumatoid arthritis, psoriasis and multiple sclerosis (4).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Reporter Cell Lines are guaranteed for 1 year from date of receipt.

FAQs for IL-17/IL-17A Reporter Cell Line (NBP2-26283). (Showing 1 - 4 of 4 FAQ).

1. I want to stain some T cell subsets (Th17 and Th1), do you have antibodies for that? I need for flow cytometer in sheep or goat.
   • Unfortunately, we have neither a CD4 antibody nor an IL-17 antibody that has been validated in sheep or goat, although if you would like to try one you would again be eligible for our Innovators Reward Program. Please contact us at innovators@novusbio.com with any questions regarding this program.
2. I'm looking to establish an ELISA for IL-17 and want your recommendation on which antibody pairs to buy. Least expensive ones, please. Also, I'd like the detection antibody to be fluorescently labeled.
   • Thank you for your patience while I reviewed our ELISA validated products for IL17. I was not sure what species you were going to be investigating, so I looked up our data for the human protein. If this is incorrect, just let me know and I can look up products for the species you will be working with. As for antibody pairs, we only had one set that have been tested together: Capture: NBP2-22531 Detection: NBP2-22532. The detection product is only available currently with a biotin conjugate. We can however offer custom conjugation for this product to any of our available conjugates. Please see the this link to see the available fluors. The typical fee for custom conjugation is usually about $80 additional. Once you know which fluor you would like, I can provide you with a quote. We do also offer several ELISA kits for this target (they typically employ HRP or Biotin detection systems, so I am not sure if they will work for your needs).
3. Our customers has ordered the HEK IL-17A Prom / LUCPorter (tm) Stable Reporter Cell Line (NBP2-26283) that is resistant to puromycin. Because he wanted to transfect the plasmideROR gamma (t) plasmid (NBP2-25279) which has the neomycin resistance, I made a range of neomycin on HEK IL-17A Prom / LUCPorter (tm) Stable Reporter Cell line. The result is surprising because even at a concentration of 4mg/ml which is normally lethal to HEK, cells are still alive. Do these cells contain another plasmid with a neomycin resistance? These are Hek 293 or T 293, or did you have another explanation?
   • The NBP2-26283 cell line was generated from using HEK293 cells from ATCC. Our QC test also confirmed that NBP2-26283 is neomycin resistance. NBP2-25279 assays with NBP2-26283 are based on transient transfection of NBP2-25279 into NBP2-26283. Hence there is no need for neomycin.
4. What is the IL-17A Prom/LUCPorter(TM) Stable Reporter Cell Line (Reference : NBP2-26283 ) promotor exact sequence? Does the Renilla Luciferase gene used has a PEST sequence for protein destabilization ?
   • The promoter contains two major ROR-response elements (RORE-1 and RORE-2). However, the promoter sequence used to generate the cell line is proprietary and we are not able to provide this informtion. The Renilla Luciferase gene used does have a PEST sequence.

Bioinformatics

• Gene Symbol: IL17A