The whole cellular lysates were prepared by harvesting the cells in 1 × cell lysis buffer and Western blotting was performed as previously described.6 , 11 (link)–13 , 18 The following antibodies were purchased from Abcam (Cambridge, MA): anti-IL-6 (ab6672, 1:500), and anti-VEGF (ab46154, 1:500). Anti-β-actin (sc-1616, 1:1000) and anti-mouse-DNMT1 (sc-52919, 1:1000) were from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-STAT3 (9139, 1:1000) and anti-phospho-STAT3 (Tyr705, 9131, 1:1000) were from Cell Signaling Technology (Danvers, Massachusetts). Anti-human-FABP4 (AF3150, 1:1000) and anti-mouse-FABP4 (AF1443, 1:500) were from R&D systems (Minneapolis, MN) and the anti-human-DNMT1 (M0231L, 1:500) was from New England Biolabs (Ipswich, MA).
Anti il 6 ab6672
Manufactured by Abcam
Sourced in United States
Anti-IL-6 (ab6672) is a lab equipment product that can be used to detect and measure the levels of the interleukin-6 (IL-6) protein in various samples. It is a reagent that can be used in immunoassays, such as ELISA, to quantify IL-6 expression.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin sc 1616, by Santa Cruz Biotechnology (2 mentions)
Anti stat3 9139, by Cell Signaling Technology (2 mentions)
Anti mouse dnmt1 sc 52919, by Santa Cruz Biotechnology (2 mentions)
Anti human fabp4 af3150, by R&D Systems (2 mentions)
Anti mouse fabp4 af1443, by R&D Systems (2 mentions)
Anti human dnmt1 m0231l, by New England Biolabs (2 mentions)
Anti vegf ab46154, by Abcam (2 mentions)
Parp 9532, by Cell Signaling Technology (1 mentions)
Gapdh mab374, by Merck Group (1 mentions)
Ciap1, by R&D Systems (1 mentions)
Iκbα sc 371, by Santa Cruz Biotechnology (1 mentions)
Nik sc 7211, by Santa Cruz Biotechnology (1 mentions)
Anti ciap2, by Abcam (1 mentions)
Nf κb2 p100 p52, by Cell Signaling Technology (1 mentions)
Wst 1 cell proliferation assay kit, by Roche (1 mentions)
Apc anti mouse cd11c clone hl3, by BD (1 mentions)
Odyssey fc system, by LI COR (1 mentions)
Trans blot turbo system, by Bio-Rad (1 mentions)
5 protocols using anti il 6 ab6672
1
Protein Expression Analysis Workflow
2
Immunoblot Analysis of Cell Signaling
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Whole-cell lysates were obtained and analyzed by immunoblot as previously described.39 (link) The following primary antibodies were used: cIAP-1 (1:2000) from R&D Systems; anti-cIAP-2 (1:1000) and anti-IL-6 (ab6672; 1:1000) from Abcam; actin (sc-1615; 1:1000), IκBα (sc-371; 1:1000), NIK (sc-7211; 1:1000) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); RIP1 (BD 610458; 1:1000) from BD Biosciences (San Jose, CA, USA); NF-κB2 p100/p52 (#4882; 1:1000) and PARP (#9532; 1:1000) from Cell Signaling Technology (Beverly, MA, USA); GAPDH (MAB374; 1:5000) from EMD Millipore (Temecula, CA, USA).
3
Protein Expression Analysis Workflow
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The whole cellular lysates were prepared by harvesting the cells in 1 × cell lysis buffer and Western blotting was performed as previously described.6 , 11 (link)–13 , 18 The following antibodies were purchased from Abcam (Cambridge, MA): anti-IL-6 (ab6672, 1:500), and anti-VEGF (ab46154, 1:500). Anti-β-actin (sc-1616, 1:1000) and anti-mouse-DNMT1 (sc-52919, 1:1000) were from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-STAT3 (9139, 1:1000) and anti-phospho-STAT3 (Tyr705, 9131, 1:1000) were from Cell Signaling Technology (Danvers, Massachusetts). Anti-human-FABP4 (AF3150, 1:1000) and anti-mouse-FABP4 (AF1443, 1:500) were from R&D systems (Minneapolis, MN) and the anti-human-DNMT1 (M0231L, 1:500) was from New England Biolabs (Ipswich, MA).
4
Molecular Profiling of STAT3 and C-Rel
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Antibodies against STAT3, phosphorylated STAT3 (p-STAT3), C-Rel, and p-C-Rel were purchased from Bioss (Woburn, MA). WST-1 cell proliferation assay kit was obtained from Roche Diagnostics (Mannheim, Germany). FITC Anti-mouse B7-1 (Clone 16-10A1), PE Anti-mouse B7-2 (Clone GL1), PE Anti-Mouse IL-12 p40/p70 (Clone C15.6) and APC Anti-mouse CD11c (Clone HL3) antibodies were purchased from BD Pharmingen (San Diego, CA). IL-6 and IL-12 ELISA kit were obtained from R&D System Inc. (Minneapolis, MN). Anti-IL-6 (ab6672) and anti-CD11c (ab33484) antibodies for immunofluorescent (IF) staining were purchased from abcam (San Francisco, CA). Anti-IL-12 (NB600-1443) antibody for IF staining was purchased from Novusbio (Littleton, CO). Mouse sIL-6R alpha Simple Step ELISA Kit (ab203360) was obtained from abcam (Cambridge, MA). Cell Fixation/Permeabilization Kits (BD554715) for intracellular cytokine analysis was purchased from BD Bioscience (San Jose, CA). Purified Rat Anti-mouse IL-6 antibody (clone MP5-20F3) was obtained from BD Biosciences. STAT3 inhibitor VI, S31-201 was purchased from Santa Cruz Biotechnology.
5
Western Blot Analysis of GCF Proteins
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40 µL of eluted GCF protein samples were separated on a 10% SDS-polyacrylamide gel and transferred to a PVDF membrane using the Trans-Blot Turbo System from Bio-Rad (Bio-Rad Laboratories, Hercules, CA, USA). The PVDF membrane was then incubated in a blocking solution (TBS 0.1% Tween-20, 5% BSA) for 1 hour. Primary antibodies were diluted in a 3% BSA solution and left to incubate overnight at 4 °C. After three washes in a 0.1% TBS-Tween-20 solution, the membrane was exposed to secondary antibodies for 2 hours at room temperature. Following another set of washes, the membranes were scanned on an Odyssey-FC system from LI-COR Biosciences. This experiment was conducted in duplicate. Primary antibodies and their respective concentrations were as follows: anti-CA1 # NBP2-76980, 1:1 000 (Novus Biological), anti-CA2 # NBP2-89522, 1:1 000 (Novus Biological), anti-MMP8 # AB56303, 1:1 000 (Abcam Inc), anti-IL-6 # AB6672, 1:500 (Abcam Inc), anti-FTH1 # 858901, 1:1 000 (BIOLEGEND).
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