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
2.Antennapedia Control peptide: 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361
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).
Read Publications using NBP2-26504 in the following applications:
Please contact technical support for detailed reconstitution instructions.
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
FIP3NF-kappa-B essential modifier
I-kappa-B kinase subunit gamma
IkB kinase gamma subunit
IkB kinase subunit gamma
IkB kinase-associated protein 1
inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase gamma
Inhibitor of nuclear factor kappa-B kinase subunit gamma
NFkappaB essential modulator
NF-kappa-B essential modulator
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.
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.