Antibodies against MMP-1 and MMP-9 were purchased from Oncogene Research Product (Cambridge, MA, USA). Antibody against caspase-3 was obtained from Calbiochem (San Diego, CA, USA). Anti-Akt, -phospho (p)-Akt (Ser 473), p-Akt (Thr 34), and -NF-κB p65 antibodies were purchased from BD PharMingen. Anti-p-NF-κB (Ser 536) antibody was obtained from Cell Signaling Technologies (Boston, MA, USA). Antibodies directed against ASM and calnexin were obtained from Abcam (Cambridge, MA, USA). Antibodies against CD55, CD71, nucleolin, p110α, poly (ADP-ribose) polymerase (PARP), and Rac1 were purchased from Santa Cruz Biotechnology. Anti-pan-cadherin, anti-PTEN, and anti-p-PTEN (Ser 380/Thr 382/Ser 385) antibodies were obtained from Thermo Fisher Scientific (New York, NY, USA). Antibody against β-actin was obtained from Sigma-Aldrich. Peroxidase-conjugated anti-mouse, -goat, and -rabbit IgG secondary antibodies were purchased from Jackson ImmunoResearch Laboratory (West Grove, PA, USA).
Nf κb p65 antibody
Manufactured by BD
Sourced in United States
The NF-κB P65 antibody is a laboratory reagent used for the detection and study of the p65 subunit of the NF-κB transcription factor. NF-κB plays a crucial role in regulating immune responses and cellular processes. This antibody can be used in various experimental techniques, such as Western blotting, immunohistochemistry, and immunoprecipitation, to identify and quantify the p65 subunit.
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Lab products found in correlation
Antibody against β actin, by Merck Group (2 mentions)
Poly adp ribose polymerase parp, by Santa Cruz Biotechnology (2 mentions)
Anti pan cadherin, by Thermo Fisher Scientific (2 mentions)
Anti pten, by Thermo Fisher Scientific (2 mentions)
Anti p nf κb ser 536 antibody, by Cell Signaling Technology (2 mentions)
Anti p pten ser 380 thr 382 ser 385, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Dmi6000b confocal microscope, by Leica (1 mentions)
Alexa fluor 555 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 647 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Formaldehyde, by Merck Group (1 mentions)
Tnf α, by BD (1 mentions)
Nis elements ar imaging software, by Nikon (1 mentions)
Lipopolysaccharides (lps), by Thermo Fisher Scientific (1 mentions)
Lysis buffer, by BD (1 mentions)
Metformin, by Thermo Fisher Scientific (1 mentions)
Ldh activity assay kit, by Merck Group (1 mentions)
5 protocols using nf κb p65 antibody
1
Antibody Characterization for Biomarker Analysis
2
Antibodies for Signaling Pathway Analysis
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Antibodies against MMP-1 and MMP-9 were purchased from Oncogene Research Product (Cambridge, MA, USA). Antibody against caspase-3 was obtained from Calbiochem (San Diego, CA, USA). Anti-Akt, -phospho (p)-Akt (Ser 473), and -NF-κB p65 antibodies were purchased from BD PharMingen. Anti-p-NF-κB (Ser 536) antibody was obtained from Cell Signaling Technologies (Boston, MA, USA). Antibodies directed against ASM and calnexin were obtained from Abcam (Cambridge, MA, USA). Antibodies against CD55, CD71, GRP78, nucleolin, p110α, poly (ADP-ribose) polymerase (PARP), and Rac1 were purchased from Santa Cruz Biotechnology. Anti-pan-cadherin, anti-PTEN, anti-p-PTEN (Ser 380), and anti-p-PTEN (Ser 380/Thr 382/Ser 385) antibodies were obtained from Thermo Fisher Scientific (New York, NY, USA). Antibody against β-actin was obtained from Sigma-Aldrich. Peroxidase-conjugated anti-mouse, -goat, and -rabbit IgG secondary antibodies were purchased from Jackson ImmunoResearch Laboratory (West Grove, PA, USA).
3
Jasminin Modulates NF-κB Translocation
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RAW 264.7 cells were cultured overnight at 37 °C; then, the cells were washed, recultured in fresh medium with 50 μM of jasminin, vehicle control, or combined 50 μM of jasminin and 3.0 μM BAY 11-7082. After 24 h of incubation, cells were fixed by 4% formaldehyde on ice, followed by permeabilization with 0.25% Triton X-100 (Bio-Rad, Hercules, CA, USA). Cells were then blocked with 5% BSA for 1 hour, followed by incubation with NF–κB P65 antibody (BD Bioscience, Palo Alto, CA, USA). After 12 h of incubation at 4 °C, the cells were washed and bathed with the antirabbit Cy3® secondary antibody (Abcam, Boston, MA, USA) for 1 h at 25 °C. After the second wash, the cells were stained with 5 ng/mL DAPI (Sigma-Aldrich, St. Louis, MO, USA). Pictures were captured under an DMI6000B confocal microscope (Leica Microsystems, Wetzlar, Hessen, Germany). The translocation of NF–κB in the nucleus was performed by obtaining the mean fluorescence intensity.
4
TGF-β and NF-κB Signaling in Breast Cancer Cells
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MCF-7 and MDA-MB-231 cells were seeded in the flow dish overnight at 37°C and then MCF-7 cells was treated with 4 μM SSA, 30 ng/ml TGFβ1, vehicle control (0.1% DMSO) or combination of 4 μM SSA and 30 ng/ml TGFβ1 for 12 h or pretreated with 4 μM SSA for 12h before adding 30 ng/ml TGFβ1 for 1h. MDA-MB-231 cells were treated with 4 μM SSA or same volume of 0.1% DMSO for 12 h. TNF-α (BD Bioscience) at a concentration of 25 ng/ml was added to the cells for 20 min. After fixation by 4% formaldehyde (Sigma-Aldrich) for 10 min, then the cells were permeabilized with 1% Trition X-100 (Bio-Rad) and blocked with 1% BSA before incubation with Phospho-Smad-2-(Ser465/467)/Smad-3-(Ser423/425) antibody (Cell signaling) and Smad4 antibody (EMD Millipore, Billerica, MA, USA) or NF-κB P65 antibody (BD Bioscience) at 4°C overnight. After washing with PBS, the cells were incubated with the Alexa Fluor® 555-conjugated secondary antibody (Life Technologies) and Alexa Fluor® 647-conjugated secondary antibody (Life Technologies) for 1 h at room temperature. After washing and staining with 5 ng/ml DAPI (Sigma-Aldrich), imaging was taken by using Nikon Eclipse Ti Laser Confocal Scanning Microscopy (Nikon Instruments Inc, Melville, NY, USA). The relative fluorescence intensity of NF-κB and p-Smad2/3 in the nucleus was quantitated by using NIS-Elements AR imaging software (Nikon).
5
NF-κB Activation and Translocation Assay
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Metformin, LPS and NF-κB activation-nuclear translocation assay kits were purchased from Peprotech, Inc. (Rocky Hill, NJ, USA). Phospho-NF-κBp65 (Ser536) antibody, NF-κB p65 antibody, ammonium pyrrolidinedithiocarbamate (PDTC) and lysis buffer were all acquired from BD Biosciences (Franklin Lakes, NJ, USA). The LDH activity assay kit was obtained from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). VCAM-
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