RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-29321
Key Product Details
Species
Human, Mouse, Rat, Bovine, Canine
Product Specifications
Application Notes
Inhibition of NF-kB activitiy.
Researchers can study the effect of p65 inhibitor peptide using a variety of methods. Following is a general protocol for KBM-5 cells. It may need to be optimized for different cell types.
Preincubate cells with appropriate amounts of inhibitory or control peptides for 1 hr and then treat with TNF or other NF-kB activating agents. Prepare nuclear extracts and check for the presence of NF-kB DNAbinding activity by EMSA (Figure 1). Nuclear extracts can be prepared as described by Bharati A, et al, 2003 and Takada et al, 2004 or using nuclear extraction kit.
Please refer to Takada et al, 2004 for further details on the use of this inhibitory peptide.
Researchers can study the effect of p65 inhibitor peptide using a variety of methods. Following is a general protocol for KBM-5 cells. It may need to be optimized for different cell types.
Preincubate cells with appropriate amounts of inhibitory or control peptides for 1 hr and then treat with TNF or other NF-kB activating agents. Prepare nuclear extracts and check for the presence of NF-kB DNAbinding activity by EMSA (Figure 1). Nuclear extracts can be prepared as described by Bharati A, et al, 2003 and Takada et al, 2004 or using nuclear extraction kit.
Please refer to Takada et al, 2004 for further details on the use of this inhibitory peptide.
Reactivity Notes
Broad: Peptide sequence is 100% conserved across multiple species.
Inhibitor Content
Inhibitor peptide (NBP2-31224): 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKKNGLLSGDEDFSS (p65 sequence: NGLLSGDEDFSS). Molecular weight: 3697.21
Antennapedia Control peptide (NBP2-29334): 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361
Scientific Data Images for RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set
![Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]](https://resources.bio-techne.com/images/products/RelA-NFkB-p65-[p-Ser529--p-Ser536]-Inhibitor-Peptide-Set-Western-Blot-NBP2-29321-img0015.jpg "Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]")
Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]
Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321] - Inhibits TNF-induced NF-kB activation: KBM-5 cells were incubated with various concentrations of peptides for 1 hr and treated with 0.1nM TNF for 30min. Nuclear extracts were prepared and NF-kB activation was analyzed by EMSA (Please refer toTakada et al, 2004 for further details). PTD-p65-P6 suppresses TNF-induced NF-kB activation by 25% at 100 uM and completely at 150 uM, where as the control peptide, PTD or p65-P6 without PTD did not have any effect.![Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]](https://resources.bio-techne.com/images/products/RelA-NFkB-p65-[p-Ser529--p-Ser536]-Inhibitor-Peptide-Set-Western-Blot-NBP2-29321-img0013.jpg "Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]")
Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]
Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321] - Effect of various p65-inhibitor peptides on binding of purified recombinant p65 protein to the DNA in vitro. Recombinant p65 protein (100 ng/sample) was incubated for 30 minutes with 50uM of various peptides in 0.025 ml and then assayed for DNA binding activity by EMSA (Please refer to Takada et al, 2004 (3) for further details).![Functional (Inhibition): RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]](https://resources.bio-techne.com/images/products/RelA-NFkB-p65-p-Ser529-p-Ser536-Inhibitor-Peptide-Set-Functional-Inhibition-NBP2-29321-img0012.jpg "Functional (Inhibition): RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]")
Functional (Inhibition): RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321]
Functional (Inhibition): RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321] - 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 different concentrations (0, 1, 5, 10, 25 and 50 uM) of Inhibitory Peptide (NBP2-29321) and Control Peptide (NBP2-29334) for 1 h. Cells were then stimulated with 1 ng/ml Flagellin (NBP2-25289) for 24 h. Secreted alkaline phosphatase (SEAP) was analyzed using SEAPorter Assay Kit (NBP2-25285). *p < 0.05 versus control peptide at the corresponding concentrations (Mann-Whitney U test).Formulation, Preparation, and Storage
Preparation Method
Preparation of 5 mM Stock Solutions
PBS* is added directly to the vials to prepare the stock solutions.
Note: Bring the solution to room temperature and quick spin the tubes before opening the caps. NF-kB p65(Ser529/536) Inhibitor Peptide: 1mg of DRQIKIWFQNRRMKWKKNGLLSGDEDFSS
Add 54.1 ul PBS* to the vial, to make a 5 mM stock solution. Mix by vortexing. Aliquot and store at -20C or -80C. Avoid repeated freeze thawing.
Control Peptide: 1 mg of DRQIKIWFQNRRMKWKK
Add 84.8 ul PBS* to the vial. Mix by vortexing. Aliquot and store at -20C or -80C. Avoid repeated freeze thawing.
*Recipe for 1X PBS:
1. Dissolve the following in 800ml distilled H2O.
- 8g of NaCl
- 0.2g of KCl
- 1.44g of Na2HPO4
- 0.24g of KH2PO4
2. Adjust pH to 7.5 with HCl.
3. Adjust volume to 1L with additional distilled H2O.
4. Sterilize by autoclaving
PBS* is added directly to the vials to prepare the stock solutions.
Note: Bring the solution to room temperature and quick spin the tubes before opening the caps. NF-kB p65(Ser529/536) Inhibitor Peptide: 1mg of DRQIKIWFQNRRMKWKKNGLLSGDEDFSS
Add 54.1 ul PBS* to the vial, to make a 5 mM stock solution. Mix by vortexing. Aliquot and store at -20C or -80C. Avoid repeated freeze thawing.
Control Peptide: 1 mg of DRQIKIWFQNRRMKWKK
Add 84.8 ul PBS* to the vial. Mix by vortexing. Aliquot and store at -20C or -80C. Avoid repeated freeze thawing.
*Recipe for 1X PBS:
1. Dissolve the following in 800ml distilled H2O.
- 8g of NaCl
- 0.2g of KCl
- 1.44g of Na2HPO4
- 0.24g of KH2PO4
2. Adjust pH to 7.5 with HCl.
3. Adjust volume to 1L with additional distilled H2O.
4. Sterilize by autoclaving
Concentration
Lyoph
Reconstitution Instructions
Please contact technical support for detailed reconstitution instructions.
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Background: RelA/NFkB p65
The Ser529/536 sites of p65 are phosphorylated during NF-kB activation, allowing p65 nuclear translocation. This p65 inhibitory peptide contains Ser529/536 sites that are phosphorylated during NF-kB activation, thereby blocking p65 Ser529/536 phosphorylation. The NF-kB p65(Ser529/536) Inhibitor Peptide can inhibit binding of recombinant p65 protein to the DNA in a dose dependent manner and maximum inhibition occurs at 50 uM (Figure 1). It can also inhibit TNF-induced NF-kB activation in cells (See Figure 2)
Long Name
v-rel Reticuloendotheliosis Viral Oncogene Homolog A
Alternate Names
NFkB p65, NFKB3, p65RelA, nf kb inhibitor, nf kb p65 inhibitor, nfkb inhibitor, nfkb p65 inhibitor, p65 inhibitor, rela inhibitor
Gene Symbol
RELA
Additional RelA/NFkB p65 Products
Product Documents for RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set
Product Specific Notices for RelA/NFkB p65 [p Ser529, p Ser536] 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.
Loading...
Loading...
Loading...