IKK gamma Inhibitor Peptide Set

Images

 
In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504] - NBD peptide blocks constitutive NF-kB as shown by EMSA. U266 cells were treated with 100 uM of control or NBD peptide for different time periods. Nuclear ...read more
In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504] - NBD peptide blocks constitutive NF-kB activation in human multiple myeloma cells. U266 cells were treated with 100 uM of control (A & B) or NBD peptide (C & ...read more
IKK gamma Inhibitor Peptide Set [NBP2-26504] - The IKK complex consists of IKKalpha + IKKbeta+ NEMO. NEMO (IKKgamma/NF-kB Essential MOdulator) is a scaffold protein required for the kinase activity of IKKalpha + ...read more
Functional (Inhibition): IKK gamma Inhibitor Peptide Set [NBP2-26504] - TLR5/NF-kB/SEAPorter HEK 293 (NBP2-26277) cells were plated in 96-well plates at 5 x 10^4 cells/well for 16 h. Cells were preincubated with ...read more

Product Details

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

Order Details

IKK gamma Inhibitor Peptide Set Summary

Immunogen
Functions as an IKK-Alpha/IKK-Beta decoy by binding to IKK-Gamma NBD, thereby preventing formation of the IKK complex.
Specificity
IKK-Gamma NEMO Binding Domain (NBD) inhibitor peptide contains a protein transduction (PTD) sequence (DRQIKIWFQNRRMKWKK) derived from antennapedia which renders the peptide cell permeable.

The control peptide consists of only the PTD sequence.
Content
1.IKK-gamma NEMO Binding Domain (NBD) Inhibitor Peptide: 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKKTALDWSWLQTE (IKK-gamma/NEMO binding sequence is underlined). Molecular weight: 3692

2.Antennapedia Control peptide: 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361
Gene
IKBKG

Applications/Dilutions

Application Notes
Inhibition of NF-kB activitiy in vivo and in vitro by interefering with IKK complex formation
Researchers can study the effect of NBD inhibitor peptide using a variety of methods. Quantitative readout assays include NF-kB/p65 ActivELISA Kit, Phospho-IkBalpha ActivELISA Kit and EMSA. Immunocytochemistry can also be used as a readout assay for visualizing the subcellular localization of NF-kB; activated NF-kB localizes to the nucleus, whereas NF-kB in the cytoplasm is generally considered be inactive.

U266 cells and EMSA assay are used to quality control every lot of the NBD inhibitor peptide set (Fig. 2). This protocol is written for U266, a human multiple myeloma cell line. Multiple myeloma is a B-cell malignancy, and a number of multiple myeloma cells lines, including U266, have been found to have constitutively active NF-kB. The EMSA assay shows that NBD suppressed the constitutive activation of NF-kB in U266 cells. The mmunocytochemistry data provides supporting evidence that the nuclear translocation of NF-kB was lost when the U266 cells were incubated with NBD (Fig.3).

Researchers must optimize assay methods for the NBD inhibitory peptide for different cell types. These include incubation time and amount of peptide used in an experiment. Depending on the cell types, morphology of cells may change after 2 hr of incubation with NBD peptide. For example, CHO cells become rounder in appearance after 2 hr incubation with NBD peptides. Since NF-kB is an important molecule for cell survival and proliferation, blockade of NF-kB activation by inhibiting IKK complex formation may prevent cell proliferation, which has been observed for CHO cells at 12 hrs. Researchers are advised to monitor the viability of cells for long-term incubation with the inhibitor. Use in Functional inhibition reported in scientific literature (PMID 24361600). Use in blocking/neutralizing reported in scientific literature (PMID 28166799).
Publications
Read Publications using
NBP2-26504 in the following applications:

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Concentration
LYOPH
Reconstitution Instructions
Please contact technical support for detailed reconstitution instructions.

Notes

The IKK-gamma inhibitory peptide contains a protein transduction (PTD) sequence (DRQIKIWFQNRRMKWKK) derived from antennapedia which renders the peptide cell permeable (Derossi et al, The third helix of the antennapedia homeodomain translocates through biological membranes. J Biol Chem. 269:10444-10450 (1994)].The control peptide consists of only the PTD sequence.

Alternate Names for IKK gamma Inhibitor Peptide Set

  • FIP-3IP2
  • FIP3NF-kappa-B essential modifier
  • FIP3P
  • I-kappa-B kinase subunit gamma
  • IkB kinase gamma subunit
  • IkB kinase subunit gamma
  • IkB kinase-associated protein 1
  • IKBKG
  • IKK gamma
  • IKKAP1
  • IKKG
  • IKK-gammaIPD2
  • incontinentia pigmenti
  • inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase gamma
  • Inhibitor of nuclear factor kappa-B kinase subunit gamma
  • IP
  • IP1
  • NEMO
  • NEMOAMCBX1
  • NFkappaB essential modulator
  • NF-kappa-B essential modulator

Background

IKK-Gamma binds to specific sequences on IKKAlpha and IKKBeta, forming the IKK complex. The IKK complex phosphorylates downstream signaling molecules during NF-kB activation. The IKK-Gamma inhibitory peptide contains an IKKAlpha and IKKBeta consensus binding sequence, called NEMO binding domain or NBD. When IKKAlpha and IKKBeta are bound to the IKKGamma NBD peptide, they can not bind to IKKGamma. Thus the formation and function of the IKK complex is blocked.

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.

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Publications for IKK gamma Inhibitor (NBP2-26504)(22)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 4 applications: B/N, Func, Func-INH, In-vitro.


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B/N
(4)
Func
(4)
Func-INH
(1)
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(1)
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Human
(5)
Mouse
(4)
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Showing Publications 1 - 10 of 22. Show All 22 Publications.
Publications using NBP2-26504 Applications Species
Alvarez-Carbonell D, Garcia-Mesa Y, Milne S et al. Toll-like receptor 3 activation selectively reverses HIV latency in microglial cells. Retrovirology. Feb 6 2017 [PMID: 28166799] (B/N, Human) B/N Human
Tanaka T, Imamura T, Yoneda M et al. Enhancement of active MMP release and invasive activity of lymph node metastatic tongue cancer cells by elevated signaling via the TNF-alpha-TNFR1-NF-kB pathway and a possible involvement of angiopoietin-like 4 in lung metastasis Int. J. Oncol. Aug 10 2016 [PMID: 27511626]
Garvin LM, Chen Y, Damsker JM, Rose MC. A novel dissociative steroid VBP15 reduces MUC5AC gene expression in airway epithelial cells but lacks the GRE mediated transcriptional properties of dexamethasone. Pulm Pharmacol Ther. Apr 28 2016 [PMID: 27133900] (Func, Human) Func Human
Li Y, Kinzenbaw DA, Modrick ML et al. Context-Dependent Effects of SOCS3 in Angiotensin II-Induced Vascular Dysfunction and Hypertension in Mice: Mechanisms and Role of Bone Marrow-Derived Cells. Am. J. Physiol. Heart Circ. Physiol. 2016 Apr 22 [PMID: 27106041] (Func, Mouse) Func Mouse
Esposito E, Napolitano G, Pescatore A et al. COMMD7 as a novel NEMO interacting protein involved in the termination of NF-kBsignaling J. Cell. Physiol. 2015 Jun 08 [PMID: 26060140] (Func, Mouse) Func Mouse
Zhang B, Shimada Y, Kuroyanagi J et al. In vivo selective imaging and inhibition of leukemia stem-like cells using the fluorescent carbocyanine derivative, DiOC5(3) Biomaterials 2015 Jun 01 [PMID: 25818410] (In-vitro, Func, Human)

Details:
IKK-gamma NEMO Binding Domain (NBD) Inhibitor Peptide used for functional assay in vitro on K562 cells and it was found to selectively kill K562 ALDH+ cells (IC50: ALDH+, 40.52 um; ALDH-, 62.38 um) after 24 hours of treatment (Fig. S7).
In-vitro, Func Human
Thein S, Pham A, Bayer KU et al. IKK regulates the deubiquitinase CYLD at the postsynaptic density. Biochem. Biophys. Res. Commun. 2014 Jun 10 [PMID: 24928390] (B/N, Rat)

Details:
Fig 2A: Primary brain hippocampal cell [post synaptic density fractions( PSD)] cultures. Cultures were treated with 5 uM or 20 uM of the inhibitory peptide or 20 uM of the control peptide. Readout system: WB analysis of ATP induced-CLYD phosphorylation. The results showed that 20 uM of the inhibitory, but not control, peptide inhibited CYLD phosphorylation (using a CYLD phosphospecific antibody) and that total CYLD (using an antibody that recognized total CYLD) expression was unchanged. The results suggested that phosphorylation of CYLD requires the regulatory subunit of IKKgamma in the PSD hippocampal cell culture model system.
B/N Rat
Kragh CL, Gysbers AM, Rockenstein E et al. Prodegenerative IkBalpha expression in oligodendroglial alpha-synuclein models of multiple system atrophy. Neurobiol Dis 2013 Dec 17 [PMID: 24361600] (Func-INH) Func-INH
Qin S, Taglienti M, Cai L et al. c-Met and NF-kB-dependent overexpression of Wnt7a and -7b and Pax2 promotes cystogenesis in polycystic kidney disease. J Am Soc Nephrol. 2012 Aug [PMID: 22677559]

Details:
Products cited: 1. IKK-gamma NEMO Binding Domain Inhibitory Peptide (IMG-2000/IMG-2000-5): Fig 6E (immortalized mouse kidney epithelial cell line) and Fig 6F (mouse kidney explants). The cell line and explants were treated with IMG-2000. Readout assay: RT
Matera MG, Calzetta L, Segreti A et al. Emerging drugs for chronic obstructive pulmonary disease. Expert Opin Emerg Drugs. 2012 Mar [PMID: 22335427]
Show All 22 Publications.

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Blogs on IKK gamma.

Regulating Immune Response Pathways with IKK beta
IKK beta, also known as IKK2, activates the NFkB complex by phosphorylating the NFkB inhibitor, IkBa. Several transcript variants, some protein-coding and some not, have been found for IKKB. The Nuclear Factor-kappa B (NF-kB) family of transcription f...  Read full blog post.

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Bioinformatics

Gene Symbol IKBKG