Immunogen	Functions as a p65 decoy through phosphorylation of the Ser529/536 sites on the peptide.
Modification	p Ser529, p Ser536
Specificity	The NF-kB p65(Ser529/536) inhibitory peptide also 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.
PreparationMethod	Preparation of 5 mM Stock SolutionsPBS* 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 DRQIKIWFQNRRMKWKKAdd 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 KH2PO42. Adjust pH to 7.5 with HCl.3. Adjust volume to 1L with additional distilled H2O. 4. Sterilize by autoclaving
Content	Inhibitor peptide (NBP2-31224): 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKKNGLLSGDEDFSS (p65 sequence: NGLLSGDEDFSS). Molecular weight: 3697.21Antennapedia Control peptide (NBP2-29334): 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361
Gene	RELA

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.

Publications

ReadPublications using NBP2-29321 in the following applications: B/N 1 publication In vitro 1 publication

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