IKK gamma Inhibitor Peptide Set
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-26504
![Functional (Inhibition): IKK gamma Inhibitor Peptide Set [NBP2-26504]](https://resources.bio-techne.com/images/products/IKK-gamma-Inhibitor-Peptide-Set-Functional-Inhibition-NBP2-26504-img0016.jpg "Functional (Inhibition): IKK gamma Inhibitor Peptide Set [NBP2-26504]")
Key Product Details
Species
Human, Mouse, Rat
Applications
Block/Neutralize, Functional Assay, Inhibition Activity
Product Specifications
Specificity
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.
Applications
Block/Neutralize (reported in scientific literature (PMID 28166799))
Functional (Inhibition) (reported in scientific literature (PMID 24361600))
Functional (Inhibition) (reported in scientific literature (PMID 24361600))
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.
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.
Inhibitor 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
2.Antennapedia Control peptide: 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361
Scientific Data Images for IKK gamma Inhibitor Peptide Set
![In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504]](https://resources.bio-techne.com/images/products/IKK-gamma-Inhibitor-Peptide-Set-In-vivo-assay-NBP2-26504-img0010.jpg "In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504]")
In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504]
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 extracts were isolated and checked for NF-kB-DNA binding activity.![In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504]](https://resources.bio-techne.com/images/products/IKK-gamma-Inhibitor-Peptide-Set-In-vivo-assay-NBP2-26504-img0012.jpg "In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504]")
In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504]
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 & D) for 12 hr, cytospun, plated on glass slides, air dried for 1 hr at room temperature and fixed with cold acetone. Slides were blocked with 5% normal goat serum for 1 hr and then incubated with rabbit polyclonal anti-human p65 antibody (A & C) followed by Ig-Alexa 594 second step. In control peptide treated cells, p65 translocates to nucleus (A), whereas NBD peptide prevents translocation of p65 into the nucleus (C). B & D: Nuclear staining with DNA binding dye.
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 + IKKbeta. NBD (NEMO Binding Domain) inhibits formation of the IKK complex (IKKalpha + IKKbeta+ NEMO) by binding to NEMO and blocking IKK complex formation. In this regard, NBD is considered to be an 'IKKalpha and IKKBeta decoy' Blocking IKK complex formation suppresses downstream events in the NF-kB signaling pathway that rely on the IKK complex including phosphorylation of IkBalpha, and subsequence IkBalpha ubiquination, degradation and release of NF-kB. Hence, NF-kB is retained in the cytoplasm in its inactive state.
Formulation, Preparation, and Storage
Concentration
Lyoph
Reconstitution Instructions
Please refer to the product PDF at novusbio.com for reconstitution instructions.
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store at -20C. Avoid freeze-thaw cycles.
Background: IKK gamma
Long Name
IkB Kinase gamma
Alternate Names
FIP3P, IKBKG, IKKAP1, NEMO
Gene Symbol
IKBKG
Additional IKK gamma Products
Product Documents for IKK gamma Inhibitor Peptide Set
Product Specific Notices for IKK gamma Inhibitor Peptide Set
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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