anti human leptin antibody  (R&D Systems)

Journal: Oncology reports

Article Title: A potential role for hepcidin in obesity-driven colorectal tumourigenesis.

doi: 10.3892/or.2017.6062

Figure Lengend Snippet: Figure 1. Brightfield images of adipocytes and quantification of IL-6, leptin and hepcidin in adipocyte secretome. (A) Mature human adipocytes (x4 magnification). (B) Mature human adipocytes with evidence of lipid filled vesicles (x20 magnification). (C) Immature preadipocytes (x10 magnifica- tion). (D) Fold change in the secreted levels of IL-6, leptin and hepcidin in DAS compared to PAS, the latter normalised to 1. Error bars denote ± SEM with * indicating statistical significance (p<0.05, n=3). DAS, differentiated adipocyte secretome; PAS, predifferentiated adipocyte secretome.

Article Snippet: For experiments involving the use of adipocyte conditioned medium upon the SW480 cells reaching 70% confluency, the cells were challenged with either the DAS or with the control (PAS) in the presence or absence of inhibitors of leptin (anti-human leptin/OB antibody MAB398; R&D Systems, Abingdon, UK) used at a final concentration of 3.2 ng/ ml; IL-6 (anti-human IL-6 antibody 500-P26G; Peprotech, London, UK) used at a final concentration of 0.1 ng/ml; or hepcidin (hepcidin antagonist peptide) as previously described by Kaplan et al (27) (RR-FDCITTGYAYTqGLSGSILS-RR; Alta Biosciences, Birmingham, UK) used at a concentration of 1 μM and the effect on cellular viability (MTT assay) and proliferation (BrdU assay) was determined, as detailed below.

Techniques:

Journal: Oncology reports

Article Title: A potential role for hepcidin in obesity-driven colorectal tumourigenesis.

doi: 10.3892/or.2017.6062

Figure Lengend Snippet: Figure 2. Cell viability and proliferation of colorectal carcinoma SW480 cells challenged with either DAS, ± leptin, IL-6 or hepcidin inhibitors. Controls included cells in preadipocyte secretome. Cells were then subjected to (A) assessment forcellular viability by an MTT assay and (B) cellular pro- liferation using a BrdU assay. Error bars denote ± SEM and n=3. Statistical significance is denoted by * and ** which represent p<0.05 and p<0.005 respec- tively, when compared to the control. DAS, differentiated adipocytes secretome.

Article Snippet: For experiments involving the use of adipocyte conditioned medium upon the SW480 cells reaching 70% confluency, the cells were challenged with either the DAS or with the control (PAS) in the presence or absence of inhibitors of leptin (anti-human leptin/OB antibody MAB398; R&D Systems, Abingdon, UK) used at a final concentration of 3.2 ng/ ml; IL-6 (anti-human IL-6 antibody 500-P26G; Peprotech, London, UK) used at a final concentration of 0.1 ng/ml; or hepcidin (hepcidin antagonist peptide) as previously described by Kaplan et al (27) (RR-FDCITTGYAYTqGLSGSILS-RR; Alta Biosciences, Birmingham, UK) used at a concentration of 1 μM and the effect on cellular viability (MTT assay) and proliferation (BrdU assay) was determined, as detailed below.

Techniques: MTT Assay, BrdU Staining, Control