On day 21 post implantation, animals were euthanized, tumor tissue was processed and western blots were performed as previously described [13 (link)]. Protein concentrations were determined using the Pierce BCA protein assay kit (Thermo Scientific, Rockford, IL, USA). Primary antibodies included CA IX (1:1000; Proteintech, Chicago, IL), HIF-1α (1:500; Bioss, Woburn, MA), NF-κB (1:1000; Cell Signaling Technology), phosphorylated NF-κB (p-NF-κB; 1:1000; Cell Signaling Technology), CD31 (1:5000; Novus Biologicals, Littleton, CO), VEGF (1:1000; Santa Cruz Biotechnology, Dallas, TX), VEGF receptor 2 (VEGFR2; 1:5000; Novus Biologicals), MMP-2 (1:500; Bioss), MMP-9 (1:600; Novus Biologicals), Vimentin (1:200, Santa Cruz Biotechnology), zona occudins-1 (ZO-1; 1:200; Biorbyt, Cambridge, UK), Occludin (1:40; Biorbyt), Aquaporin-1 (1:1000; Abcam, Cambridge, UK) and Aquaporin-4 (1:1000; Abcam), and β-actin (1:6000; Abcam). Densitometry was used to determine the ratio of the target to β-actin (VisionWorks LS 7.1 software; UVP, Upland, CA).
Phosphorylated nf κb p nf κb
Manufactured by Cell Signaling Technology
Sourced in United Kingdom
Phosphorylated NF-κB (p-NF-κB) is a lab equipment product that detects the phosphorylated form of the NF-κB protein. NF-κB is a transcription factor that plays a key role in regulating immune response, inflammation, and cell survival. The phosphorylated form of NF-κB is an indicator of its activation status.
Automatically generated - may contain errors
Lab products found in correlation
Vascular endothelial growth factor (vegf), by Santa Cruz Biotechnology (1 mentions)
Aquaporin 1, by Abcam (1 mentions)
β actin, by Abcam (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Vimentin, by Santa Cruz Biotechnology (1 mentions)
Hif 1α, by Bioss Antibodies (1 mentions)
β actin, by Analytik Jena (1 mentions)
Mmp 2, by Bioss Antibodies (1 mentions)
Aquaporin 4, by Abcam (1 mentions)
Nf κb, by Cell Signaling Technology (1 mentions)
Taq polymerase, by Roche (1 mentions)
Ecl solution, by GE Healthcare (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Lamin b, by Santa Cruz Biotechnology (1 mentions)
Dulbecco s modified eagle s medium dmem, by Welgene (1 mentions)
2 protocols using phosphorylated nf κb p nf κb
1
Tumor Protein Analysis by Western Blot
2
HaCaT Cell Signaling Pathway Activation
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HaCaT cells were obtained from the cell line service (CLS, Eppelheim, Germany). Penicillin-streptomycin (P/S), fetal bovine serum (FBS) and goat serum (16210-064) were obtained from Gibco; Thermo fisher Scientific, Inc. Dulbecco's modified Eagle's medium (DMEM) was purchased form Welgene. CD4 (ab183685) and CD8 (ab209775) antibodies were obtained from Abcam. Antibodies to phosphorylated-extracellular signal-regulated kinase (p-ERK, #4370), extracellular signal-regulated kinase (ERK, #4695), phosphorylated-p38 (p-p38, #4511), p38 (#9212), phosphorylated-Jun N-terminal kinase (p-JNK, #4668), Jun N-terminal kinase (JNK, #9258) and phosphorylated-NF-κB (p-NF-κB, #3033) were purchased from Cell Signaling Technology, Inc. Lamin B was purchased from Santa Cruz Biotechnology, Inc. Protease inhibitor cocktail (#P8340) and phosphatase inhibitor cocktail (#P0044, #P5726) were purchased from Sigma-Aldrich; Merck KGaA. ECL solution (RPN2106) was obtained from GE Healthcare Life Sciences. PCR primers glyceraldehyde 3-phosphate dehydrogenase (GAPDH), TARC/CCL17, GM-CSF, CXCL-8 were obtained from Genotech. Taq polymerase was obtained from KaPa BioSystems. TNF-α (#285-IF), IFN-γ (#210-TA) were purchased from R&D Systems.
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