NFkB p50 (NLS) Inhibitor Peptide Set

Functional (Inhibition): NFkB p105/p50 Inhibitor Peptide Set [NBP2-29323] - 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 (NBP2-29323) 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).

• Reactivity: Hu, Mu, Rt, Bv, Ca, Ch, Xp
• Applications: Inhibition Activity
• Concentration: LYOPH

NFkB p50 (NLS) Inhibitor Peptide Set Summary

• Immunogen: Functions as a p50 decoy by blocking the intracellular recognition mechanism of p50 NLS.
• Specificity: The p50 NLS 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
• 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. p50 (NLS)
  Inhibitor Peptide: 1 mg of DRQIKIWFQNRRMKWKKVQRKRQKLM
  Add 57 ul of 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
• Content: NF-kB p50 (NLS) Inhibitor peptide: 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKKVQRKRQKLM (p50 NLS sequence: VQRKRQKLM). Molecular weight: 3529. Antennapedia Control peptide: 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361
• Gene: NFKB1

Applications/Dilutions

• Dilutions:
  • Functional (Inhibition) reported in scientific literature (PMID 24384074)
• Application Notes: Inhibition of NF-kB p50(NLS) activity. The inhibitor peptide is designed to block p50 nuclear translocation. Optimal peptide concentrations and incubation times vary between model systems and should be determined empirically by users. A 100 uM final concentration may be a useful starting point. Please refer to Lin et al (1995) for additional information about how the inhibitor peptide has been used to block p50 nuclear translocation.

Reactivity Notes

Broad: Peptide sequence is 100% conserved across multiple species.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Concentration: LYOPH
• Reconstitution Instructions: Please contact technical support for detailed reconstitution instructions.

Alternate Names for NFkB p50 (NLS) Inhibitor Peptide Set

• CVID12
• EBP-1
• KBF1
• NFkappaB
• NF-kappaB
• NF-kappa-B1
• NFKB1
• NF-kB1
• NFKB-p105
• NFKB-p50
• nuclear factor kappa B subunit 1
• p105
• P50

Background

The NLS sequence is required for nuclear translocation of NF-kB p50. In stimulated cells, NF-kB is in the cytoplasm in an inactive form and the NLS is masked. The degradation of IkBalpha during NF-kB signaling unmasks the NLS of NF-kB subunits and NF-kB is translocated to the nucleus. The p50 inhibitory peptide contains the p50 NLS sequence (VQRKRQKLM) and blocks the intracellular recognition mechanism for the NLS present on p50. Consequently p50 nuclear translocation is blocked.

Limitations

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.

Publications for NFkB p50 (NLS) Inhibitor (NBP2-29323)(12)

Publications using NBP2-29323

• Khan, A;Bakhru, P;Saikolappan, S;Das, K;Soudani, E;Singh, CR;Estrella, JL;Zhang, D;Pasare, C;Ma, Y;Sun, J;Wang, J;Hunter, RL;Tony Eissa, N;Dhandayuthapani, S;Jagannath, C; An autophagy-inducing and TLR-2 activating BCG vaccine induces a robust protection against tuberculosis in mice NPJ Vaccines 2019-08-05 [PMID: 31396406] (B/N, Mouse)
• Karim ZA, Hensch NR, Qasim H et al. Role of I kappa B Kinase beta in Regulating the Remodeling of the CARMA1-Bcl10-MALT1 Complex Biochem Biophys Res Commun. 2018-06-01 [PMID: 29649481] (Mouse)
• Ishizuka EK, Filgueiras LR, Rios FJ et al. PAFR activation of NF-kB p65 or p105 precursor dictates pro- and anti-inflammatory responses during TLR activation in murine macrophages. Sci Rep. 2016-08-24 [PMID: 27554194] (Mouse)
• Stassi Stefano, Cauda Valentina, Canavese Giancarlo, Pirri Candido Fabrizio. Flexible tactile sensing based on piezoresistive composites: a review. Sensors (Basel). 2014-01-01 [PMID: 24638126] (Human)
• Bakhru P, Sirisaengtaksin N, Soudani E et al. BCG vaccine mediated reduction in the MHC-II expression of macrophages and dendritic cells is reversed by activation of Toll-like receptors 7 and 9. Cell Immunol 2014-01-01 [PMID: 24384074] (Forced Spectroscopy, Mouse)
• Park Jang-Yeon, Moeller Steen, Goerke Ute et al. Short echo-time 3D radial gradient-echo MRI using concurrent dephasing and excitation. Magn Reson Med. 2012-02-01 [PMID: 21702064] (Human)
• Ababneh Sufyan Y, Prescott Jeff W, Gurcan Metin N. Automatic graph-cut based segmentation of bones from knee magnetic resonance images for osteoarthritis research. Med Image Anal. 2011-08-01 [PMID: 21474362] (Human)
• Branski RC, Zhou H, Sandulache VC et al. Cyclooxygenase-2 signaling in vocal fold fibroblasts. Laryngoscope. 2010-09-01 [PMID: 20717945]
  
  Details:
  Product cited: 1. NF-kB p50 (NLS) Inhibitory Peptide Set (IMG-2004): HVOX human vocal fold fibroblasts cell line, Fig 3A, B. Readout assays measuring ratio of nuclear to cytoplasmic p50 (Fig 3A) and COX-2 transcription (Fig 3B) were used to measure the ef
• Fang X, Neyrinck AP, Matthay MA, Lee JW. Allogeneic human mesenchymal stem cells restore epithelial protein permeability in cultured human alveolar type II cells by secretion of angiopoietin-1. J Biol Chem. 2010-08-20 [PMID: 20554518]
  
  Details:
  Product cited: 1. NF-kB p50 (NLS) Inhibitory Peptide Set (IMG-2004): Primary human alveolar epithelial Type II Cells, Fig 9D. An epithelial permeability to protein assay was used as a readout to measure the effects of the p50 peptide. The p50 peptide was
• Mohamed JS, Lopez MA, Cox GA, Boriek AM. Anisotropic regulation of Ankrd2 gene expression in skeletal muscle by mechanical stretch. FASEB J. 2010-09-01 [PMID: 20442316]
  
  Details:
  1. NF-kB p50 (NLS) Inhibitory Peptide Set (IMG-2004): Fig 4AB.
• Buchau AS, MacLeod DT, Morizane S et al. Bcl-3 acts as an innate immune modulator by controlling antimicrobial responses in keratinocytes. J Invest Dermatol. 2009-09-01 [PMID: 19282837]
• Haker Steven, Wells William M 3rd, Warfield Simon K et al. Combining classifiers using their receiver operating characteristics and maximum likelihood estimation. Med Image Comput Comput Assist Interv. 2005-01-01 [PMID: 16685884]

FAQs for NFkB p50 (NLS) Inhibitor (NBP2-29323). (Showing 1 - 1 of 1 FAQs).

1. Please let us know the buffer composition prior to lyophilization. Is it lyophilized from H2O?
   • Thank you for your inquiry about NBP2-20323, the NFkB p105/p50 Inhibitor Peptide Set. This product is lyophilized from a PBS buffer.

Bioinformatics

• Gene Symbol: NFKB1