TLR4 Inhibitor Peptide Set

Images

 
TLR4 Inhibitor Peptide Set [NBP2-26244] - The inhibitor peptide inhibits TLR4 signaling by blocking interactions between TLR4 and its adaptors Mal/TIRAP and TRAM.
Blocking/Neutralizing: TLR4 Inhibitor Peptide Set [NBP2-26244] - iBMDM cultures were pretreated with 5uM of inhibitor or control prior to stimulation with 20 ng/ml LPS. Negative (LPS-, LPS-/CP7+, LPS-/CP7+) and positive ...read more
Blocking/Neutralizing: TLR4 Inhibitor Peptide Set [NBP2-26244] - TLR4/MD-2/CD14/NF-kB/SEAPorter stably transfected cells were plated in 96-well plates at 1x10^5 cells/well. After 16 h, cells were preincubated with ...read more

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications Flow, Func, In vitro, B/N
Concentration
LYOPH

Order Details

TLR4 Inhibitor Peptide Set Summary

Immunogen
TLR4 inhibitor and control peptides are quality controlled in vitro using the TLR4/MD-2/CD14/NF-kB/SEAP cell line with SEAP as a readout assay (Fig 3).
Specificity
TLR4 inhibitor and control peptides are quality controlled in vitro using the TLR4/MD-2/CD14/NF-kB/SEAP cell line with SEAP as a readout assay (Fig 3).
Preparation
Method
Preparation of 5 mM VIPER and CP7 Stock Solutions
Note: Bring the peptides to room temperature and quick spin the tubes before opening the caps.

VIPER: A final volume of 72 ul will make a 5 mM stock solution. Add 72 ul sterile H20 to the tube of peptide. Carefully pipet to ensure all of the peptide is dissolved.

CP7: A final volume of 76 ul will make a 5 mM stock solution. Add 76 ul sterile H20 to the tube of peptide. Carefully pipet to ensure all of the peptide is dissolved.

The stock solutions may be diluted further to make working solutions. Dilute according to the needs for your assay. For example dilute 5 mM stock solutions 1:10 in sterile 1X PBS or cell culture media to make 500 uM working solutions. Working solutions should be made fresh daily and not stored.
Content
VIPER: A TLR4 Inhibitory Peptide: 1 mg (lyophilized) KYSFKLILAEYRRRRRRRRR (VIPER sequence: KYSFKLILAEY). Molecular weight: 2780.3

CP7, Control Peptide: 1 mg (lyophilized) RNTISGNIYSARRRRRRRRR (Control sequence: RNTISGNIYSA). Molecular weight: 2601
Gene
TLR4

Applications/Dilutions

Application Notes
The inhibitor is used in assays to inhibit TLR4 activation; see Figure 3 and also refer to Lysakova-Devine et al (2010) for examples. Optimal inhibitor concentrations should be established through titration and may vary between model systems. We recommend an initial titration from 0-30 uM for in vitro assays (Fig 3). Control concentrations should mirror inhibitor concentrations. Inhibitor and control should be preincubated with cells prior to ligand activation to allow sufficient time for the peptides to enter from the media into the cell. We typically preincubate with inhibitor and control peptides for 2 h prior to TLR4 activation with LPS (Fig 3); however, optimal preincubation times may vary between model systems.

The TLR4/MD-2/CD14 stably transfected cell line is a useful positive control model system for studying inhibition of TLR4 activation by VIPER (Fig 3). SEAP is used as a readout assay in Figure 3 to measure TLR4 inhibition.

A novel model system is shown in Figure 1 where TLR4 inhibitor peptide, but not CP7, inhibited TLR4 activation in Mal-deficient immortalized mouse bone marrow-derived macrophages (iBMDMs). In these iBMDMs, the inhibitor targets TLR4-TRAM, but not TLR-Mal, interactions as Mal is not expressed. TNF-alpha is used as a readout assay in Figure 1 to measure inhibition. Use in Function reported in scientific literature (PMID 24630524) Use in blocking / neutralizing reported in multiple pieces of scientific literature FLOW application reported by Alvarez et al - Conference on Retroviruses and Opportunistic Infections - CROI 2014 (Abstract: The NCOR2-Nurr1-CoREST Transrepression Axis Impairs HIV Reactivation in Latently Infected Microglial Cells) Use in Binding inhibition reported in scientific literature (PMID 27897392).
Reviewed Applications
Read 1 Review rated 5
using
NBP2-26244 in the following application:

Publications
Read Publications using
NBP2-26244 in the following applications:

Reactivity Notes

Human and Mouse. Rat reactivity reported in scientific literature (PMID: 25811788)

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
Form : White Solid
Purity : >95% by HPLC
Concentration
LYOPH
Reconstitution Instructions
Please contact technical support for detailed reconstitution instructions.

Notes

As applications and experimental paradigms vary, each researcher should determine the suitability of VIPER for their individual model systems. Likewise, each researcher should determine optimal concentrations, readout assays and other relevant parameters such as time courses for their model systems. Signal transduction pathways and model systems are complex, as such results may vary from examples shown herein or described in the published literature.

Alternate Names for TLR4 Inhibitor Peptide Set

  • ARMD10
  • CD284 antigen
  • CD284
  • hTollhomolog of Drosophila toll
  • TLR4
  • TOLL
  • toll-like receptor 4

Background

VIPER is a TLR4 specific inhibitory peptide that potently inhibits TLR4-mediated responses and is inert towards other TLR pathways (Bowie et al, 2010; Lysakova-Devine et al, 2010). The TLR4 inhibitor consists of an 11 amino acid (KYSFKLILAEY) inhibitory sequence, derived from the A46 vaccinia virus protein, linked to a 9R (RRRRRRRRR) homopolymer delivery sequence.

CP7 is an inert control peptide. The control consists of an 11 amino acid (RNTISGNIYSA) inert sequence also linked to a 9R homopolymer delivery sequence. The 9R sequence efficiently delivers the peptides into cells. CP7 is designed to be a negative control peptide for inhibitory assays.

We note that this inhibitor is the first viral inhibitory peptide identified that is specific for a particular TLR pathway. Inhibition of TLR4 within the context of the overall TLR signaling network is illustrated in Figure 2. VIPER should be useful for elucidating new mechanisms and molecular interactions involved in TLR4 signaling, and may also have potential for leading to the development of a new class of novel therapeutics that perturb TLR4 signaling by targeting Mal and TRAM TIR domains. The identification of additional short virally derived peptides that perturb TLR signaling is sure to open a new era in knowledge about how viruses specifically interact with host immune systems.

Limitations

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

Customers Who Viewed This Item Also Viewed...

NB100-56722
Species: Hu, Mu, Rt, Ca, Rb
Applications: WB, DB, ELISA, Flow, Func, ICC/IF, IP, In vitro, B/N, Flow-CS
7270-IL/CF
Species: Hu
NB100-56655
Species: Hu, Mu, Rt
Applications: WB, IHC, IHC-P
NBP2-29328
Species: Hu, Mu, Rt, Xp, Ye, Ze
Applications: ELISA
NB100-56583
Species: Hu, Rt, Rb
Applications: WB, Flow, IHC, IHC-P, KO
NBP2-24729
Species: Hu, Mu, Rt, Ca, Eq, Pm, Pm
Applications: WB, Simple Western, DB, ELISA, Flow, Func, ICC/IF, IHC, IHC-P, IP, In vitro, B/N, CyTOF-ready, Flow-IC, KD
NBP2-24875
Species: Hu, Mu, Ca
Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP, BA, B/N, Flow-IC
NBP2-67360
Species: Hu, Mu, Rt
Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP
MAB208
Species: Hu
Applications: WB, Simple Western, CyTOF-ready, ELISA(Cap), ELISA(Det), ICC, ICFlow, Neut, ELISA(Sta)
NB120-13810
Species: Hu, Mu, Rt, Po
Applications: WB, Simple Western, IB, ICC/IF, IP
NB100-524
Species: Hu, Mu
Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, Flow-IC
NBP2-24787
Species: Hu, Mu, Ca
Applications: WB, DB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, Flow-CS, Flow-IC
NB100-2322
Species: Hu, Mu, Rt, Bv, Ca, Rb, Sh
Applications: WB, Simple Western, ELISA, Flow, ICC/IF, IHC, IHC-P, Flow-IC, KD
Supplier Logo

Publications for TLR4 Inhibitor (NBP2-26244)(42)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Mouse;Human.

We have publications tested in 9 applications: B/N, BindInhib, FLOW, Func, Func-Inh, In vitro, In vivo, In-vitro, In-vivo.


Filter By Application
B/N
(11)
BindInhib
(1)
FLOW
(1)
Func
(13)
Func-Inh
(1)
In vitro
(7)
In vivo
(1)
In-vitro
(1)
In-vivo
(1)
All Applications
Filter By Species
Human
(14)
Mouse
(11)
Rat
(2)
Mouse;Human
(1)
All Species
Showing Publications 1 - 10 of 42. Show All 42 Publications.
Publications using NBP2-26244 Applications Species
Chehimi M, Ward R, Pestel J et al. Omega-3 Polyunsaturated Fatty Acids Inhibit IL-17A Secretion through Decreased ICAM-1 Expression in T Cells Co-Cultured with Adipose-Derived Stem Cells Harvested from Adipose Tissues of Obese Subjects Mol Nutr Food Res Mar 8 2019 [PMID: 30848861] (B/N, Human) B/N Human
Tun X, Yasukawa K, Yamada KI. Nitric Oxide Is Involved in Activation of Toll-Like Receptor 4 Signaling through Tyrosine Nitration of Src Homology Protein Tyrosine Phosphatase 2 in Murine Dextran Sulfate-Induced Colitis Biol. Pharm. Bull. Dec 4 2018 [PMID: 30504685] (B/N, Mouse) B/N Mouse
Estruch M, Sanchez-Quesada JL, Ordonez-Llanos J, Benitez S. Inflammatory intracellular pathways activated by electronegative LDL in monocytes. Biochim. Biophys. Acta. May 25 2016 [PMID: 27235719]
Mizuno Y, Taguchi T Self-assembled dodecyl group-modified gelatin microparticle-based hydrogels with angiogenic properties NPG Asia Mater Dec 1 2020
Li B, Yan C, Wu J et al. Clonorchis sinensis ESPs enhance the activation of hepatic stellate cells by a cross-talk of TLR4 and TGF-beta/Smads Signaling pathway Acta Trop. Dec 17 2019 [PMID: 31862462] (Func) Func
Harada Y, Zhang J, Imari K et al. Cathepsin E in neutrophils contributes to the generation of neuropathic pain in experimental autoimmune encephalomyelitis Pain 2019 Sep 01 [PMID: 31095099] (Mouse) Mouse
Nishida M, Saegusa J, Tanaka S, Morinobu A. S100A12 facilitates osteoclast differentiation from human monocytes. PLoS ONE Sep 20 2018 [PMID: 30235276] (Human) Human
Sahoo SS, Pratheek BM, Meena VS et al. VIPER regulates naive T cell activation and effector responses: Implication in TLR4 associated acute stage T cell responses Sci Rep May 8 2018 [PMID: 29740052] (B/N, Mouse) B/N Mouse
Li Q, Dai C, Xue R et al. S100A4 Protects Myeloid-Derived Suppressor Cells from Intrinsic ApoptosisviaTLR4-ERK1/2 Signaling. Front Immunol. Mar 5 2018 [PMID: 29556233] (B/N) B/N
Inoue H, Shirakawa J, Togashi Y et al. Signaling between pancreatic B-cells and macrophages via S100 calcium-binding protein A8 exacerbates B-cell apoptosis and islet inflammation. J. Biol. Chem. Mar 1 2018 [PMID: 29496993] (B/N) B/N
Show All 42 Publications.

Review for TLR4 Inhibitor (NBP2-26244) (1) 51

Average Rating: 5
(Based on 1 review)
We have 1 review tested in 1 species: Human.

Reviews using NBP2-26244:
Filter by Applications
ELISA
(1)
All Applications
Filter by Species
Human
(1)
All Species
Images Ratings Applications Species Date Details
ELISA TLR4 NBP2-26244
Enlarge
reviewed by:
anonymous aa
ELISA Human 09/15/2014
View

Summary

ApplicationELISA
Sample TestedTHP-1 cell culture supernatant
SpeciesHuman
CommentsLPS induced TNF alpha release was blocked by Viper peptide.

Comments

CommentsLPS induced TNF alpha release was blocked by Viper peptide.

Product General Protocols

View specific protocols for TLR4 Inhibitor (NBP2-26244): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for TLR4 Inhibitor (NBP2-26244) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Other Available Formats

NBP2-26244

Additional TLR4 Products

Bioinformatics Tool for TLR4 Inhibitor (NBP2-26244)

Discover related pathways, diseases and genes to TLR4 Inhibitor (NBP2-26244). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for TLR4 Inhibitor (NBP2-26244)

Discover more about diseases related to TLR4 Inhibitor (NBP2-26244).
 

Pathways for TLR4 Inhibitor (NBP2-26244)

View related products by pathway.

PTMs for TLR4 Inhibitor (NBP2-26244)

Learn more about PTMs related to TLR4 Inhibitor (NBP2-26244).
 

Research Areas for TLR4 Inhibitor (NBP2-26244)

Find related products by research area.

Blogs on TLR4. Showing 1-10 of 11 blog posts - Show all blog posts.

PAMPs and DAMPs: What is the same and what is different about these molecules?
By Victoria OsinskiWhat are PAMPs and DAMPsInflammation results from stimuli signaling damage or infection. The inflammatory response can be beneficial or harmful depending on the type and duration of stimuli. The s...  Read full blog post.

How to identify B cell subsets using flow cytometry
By Victoria OsinskiUsing flow cytometry to identify B cell subsetsIdentifying cellular subsets by flow cytometry requires careful and thorough planning in order to ensure the correct subset of cells are identified a...  Read full blog post.

Lipopolysaccharide from gut microbiome localizes in human atherosclerotic plaques and promotes TLR4-mediated oxidative stress
By Jamshed Arslan, Pharm. D., PhD. Atherosclerosis is a chronic inflammatory condition in which plaques of fats and other substances slowly buildup on the inner walls of arteries to restrict blood flow. In atheroscle...  Read full blog post.

Toll-like receptors in the intestinal epithelial cells
By Jamshed Arslan, Pharm. D., PhD. Toll-like receptors (TLRs) are microbe-sensing proteins that act as first responders to danger signals. TLRs help the intestinal epithelial cells (IECs) recognize commensal bacteria...  Read full blog post.

The role of STING/TMEM173 in gamma and encephalitis Herpes Simplex Virus (HSV)
Stimulator of interferon genes (STING), also known as TMEM173, promotes the production of the interferon’s IFN-alpha and IFN-beta.  STING possesses three functional domains: a cytoplasmic C-terminal tail, a central globular domain, and four N-...  Read full blog post.

TRIF/TICAM1 and mitochondrial dynamics in the innate immune response
TRIF, also known as toll like receptor adaptor molecule 1 or TICAM1, is known for its role in invading foreign pathogens as part of our innate immune response. TRIF/TICAM1 is a TIR-domain adaptor protein (toll/interleukin-1 receptor) that interacts...  Read full blog post.

The role of TLR4 in breast cancer
Toll like receptors (TLRs) are highly conserved proteins that are first known for their role in pathogen recognition and immune response activation.  In order to elicit the necessary immune response in reaction to a foreign pathogen, TLRs trigger cy...  Read full blog post.

cIAP2 - balancing cell death and cell survival
The inhibitor of apoptosis proteins (IAPs) are important regulators of cell death and inflammation. The cellular inhibitor of apoptosis protein 2 (cIAP2) contains three Baculovirus IAP repeat (BIR) domains, a Ubiquitin associated (UBA) domain, and...  Read full blog post.

TLR4 - A Guardian of Innate Immunity
Toll-like receptor 4 (TLR4) belongs to the family of Toll-like receptors (TLR), and plays a main role in pathogen recognition and innate immunity system activation. The TLR family members are highly conserved proteins that all contain a high degree of...  Read full blog post.

IRAK4: The "master IRAK" critical for initiating immune responses
IRAK4, also known as Interleukin-1 receptor-associated kinase 4, is a serine/threonine-protein kinase that plays a critical role in initiating innate and adaptive immune responses against foreign pathogens. It activates NF-kappaB in both Toll-like rec...  Read full blog post.

Showing 1-10 of 11 blog posts - Show all blog posts.

Customers Who Bought This Also Bought

Contact Information

Product PDFs

Recent Reviews

5
5
1
4
0
3
0
2
0
1
0

anonymous aa
09/15/2014
Application: ELISA
Species: Human

Bioinformatics

Gene Symbol TLR4