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![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).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Inhibitor-Peptide-Set-Blocking-Neutralizing-NBP2-26505-img0008.jpg "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).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Inhibitor-Peptide-Set-Blocking-Neutralizing-NBP2-26505-img0009.jpg)
![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).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-p-Ser276-Inhibitor-Peptide-Set-Functional-Inhibition-NBP2-26505-img0010.jpg)
![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.](http://images.novusbio.com/fullsize/IKK-gamma-Inhibitor-Peptide-Set-In-vivo-assay-NBP2-26504-img0010.jpg "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] - 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.](http://images.novusbio.com/fullsize/IKK-gamma-Inhibitor-Peptide-Set-In-vivo-assay-NBP2-26504-img0012.jpg)
![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.](http://images.novusbio.com/fullsize/IKK-gamma-Inhibitor-Peptide-Set-NBP2-26504-img0014.jpg)
![Functional (Inhibition): IKK gamma Inhibitor Peptide Set [NBP2-26504] - 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 (NBP2-26504) 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).](http://images.novusbio.com/fullsize/IKK-gamma-Inhibitor-Peptide-Set-Functional-Inhibition-NBP2-26504-img0016.jpg)
![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).](http://images.novusbio.com/fullsize/NFkB-p105-p50-Inhibitor-Peptide-Set-Functional-Inhibition-NBP2-29323-img0001.jpg "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).")
![Western Blot: TNF-alpha Antibody (6N1E7) [NBP2-27223] - Analysis using Azide Free version of NBP2-27223. TNF alpha recombinant protein using TNF alpha antibody at 3 ug/mL. Goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used for this test.](http://images.novusbio.com/fullsize/TNF-alpha-Antibody-6N1E7-Western-Blot-NBP2-27223-img0004.jpg "Western Blot: TNF-alpha Antibody (6N1E7) [NBP2-27223] - Analysis using Azide Free version of NBP2-27223. TNF alpha recombinant protein using TNF alpha antibody at 3 ug/mL. Goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used for this test.")
![<b>Biological Strategies Validation. </b>Flow Cytometry: TNF-alpha Antibody (6N1E7) [NBP2-27223] - Human PBMC were stimulated with PMA (10 ng/mL) and Ionomycin (500 ng/mL) for 5h in presence of Brefeldin A. At the end of stimulation, cells were surface stained before testing by intracellular staining. Fixed and permeabilized cells were incubated with 10 uL (0.5 ug) of TNF- alpha antibody for 30 minutes on ice. Lymphocyte population was gated for analysis to show TNF alpha positive T cells.](http://images.novusbio.com/fullsize/TNF-alpha-Antibody-6N1E7-Flow-Cytometry-NBP2-27223-img0002.jpg)
![Inhibition Activity: TNF-alpha Antibody (6N1E7) [NBP2-27223] - Validation of inhibitory effects of TNF alpha antibody on TNF alpha activity in the cell line. NF-kB/SEAPorter cells were plated in 96-well plate at 5 x 10^4 cells per well for 16 h. Cells were pre-incubated with TNF alpha antibody at 10 and 20 ug/ml for 1 hr, then treated with 10 ng/ml TNF alpha or 10 ng/ml PMA. After 24h, the secreted alkaline phosphatase (SEAP) was analyzed using the SEAPorterAssay Kit . Image using the Azide Free format of this antibody.](http://images.novusbio.com/fullsize/TNF-alpha-Antibody-6N1E7-Inhibition-Activity-NBP2-27223-img0006.jpg)
