RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-26505
Key Product Details
Species
Human, Mouse, Rat, Bovine, Canine, Mammal
Applications
Block/Neutralize, Functional Assay, Inhibition Activity
Product Specifications
Applications
Functional (Inhibition) (reported in scientific literature (PMID 24737129))
Application Notes
Application: Inhibition of DNA binding activity of NF-kB.
Researchers can study the effect of p65 inhibitor peptide using a variety of methods. This is a general protocol and may need to be optimized. 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 DNA-binding activity by EMSA (Figure 1). Nuclear extracts can be prepared as described by Bharati A, et al. and Takada et al 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. This is a general protocol and may need to be optimized. 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 DNA-binding activity by EMSA (Figure 1). Nuclear extracts can be prepared as described by Bharati A, et al. and Takada et al 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
NF-kB p65 (Ser276) Inhibitor peptide (NBP2-31221): 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKKQLRRPSDRELSE (p65 sequence: QLRRPSDRELSE). Molecular weight: 3830
Antennapedia Control peptide (NBP2-29334): 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361
Antennapedia Control peptide (NBP2-29334): 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361
Scientific Data Images for RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set
Blocking/Neutralizing: RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505] - 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 for further details).
Blocking/Neutralizing: RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505] - PTD-p65-P1 peptide 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 30 min. Nuclear extracts were prepared and NF-kB activation was analyzed by EMSA (Please refer to Takada et al, 2004 for further details).
![Functional (Inhibition): RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505]](https://resources.bio-techne.com/images/products/RelA-NFkB-p65-p-Ser276-Inhibitor-Peptide-Set-Functional-Inhibition-NBP2-26505-img0010.jpg "Functional (Inhibition): RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505]")
Functional (Inhibition): RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505]
Functional (Inhibition): RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505] - 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 (NBP2-26505) 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 (Ser276) Inhibitor Peptide: 1 mg of DRQIKIWFQNRRMKWKKQLRRPSDRELSE)
Add 52.2 ul of PBS* to the vial to make a 5 mM stock solution. Mix by vortexing. Aliquot and store at -20 degree C or -80 degree C. 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 20 degree C or -80 degree C. 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 (Ser276) Inhibitor Peptide: 1 mg of DRQIKIWFQNRRMKWKKQLRRPSDRELSE)
Add 52.2 ul of PBS* to the vial to make a 5 mM stock solution. Mix by vortexing. Aliquot and store at -20 degree C or -80 degree C. 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 20 degree C or -80 degree C. 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
Store at -20C. Avoid freeze-thaw cycles.
Background: RelA/NFkB p65
The Ser276 site of p65 is phosphorylated during NF-kB activation, allowing p65 nuclear translocation. This p65 inhibitory peptide contains a Ser276 site that is phosphorylated during NF-kB activation, thereby blocking p65 Ser276 phosphorylation (Bharti et al, 2003; Takada et al, 2004). The NF-kB p65 (Ser276), also known as PTD-p65-P1, inhibitory peptide also contains a protein transduction (PTD) sequence (DRQIKIWFQNRRMKWKK) derived from antennapedia which renders the peptide cell permeable (Derossi et al, 1994). The control peptide consists of only the PTD sequence. The PTD-p65-P1 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 vivo. PTD-p65-P1 suppresses TNF-induced NF-kB activation by 25% at 100 uM and completely at 150 uM (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 Ser276] Inhibitor Peptide Set
Product Specific Notices for RelA/NFkB p65 [p Ser276] 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...
Loading...