Human/Primate Angiopoietin-like Protein 4/ANGPTL4 Antibody

Detection of Human/Primate Angiopoietin‑like Protein 4/ANGPTL4 by Simple Western^TM. Simple Western lane view shows lysates of Human Diabetic Adipose, loaded at 0.2 mg/mL. A specific band was detected for Angiopoietin‑like Protein 4/ANGPTL4 at approximately 59 kDa (as indicated) using 50 µg/mL of Goat Anti-Human/Primate Angiopoietin‑like Protein 4/ANGPTL4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3485). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Angiopoietin-like Protein 4/ANGPTL4 by Western Blot NiCl2 activates the expression of ANGPTL4 and HIF1-alpha in BEAS-2B cells. (A) ANGPTL4 protein expression of BEAS-2B cells exposed to NiCl2 (0, 0.06, 0.12, and 0.25 mM) for 24 h was performed using western blotting. (B) The mRNA level of cells exposed to NiCl2 for 6 h was performed by real time-PCR. Significant differences from the untreated cells are indicated by ** p < 0.01 or *** p < 0.001 (C,D) Time-course analysis was also performed on BEAS-2B cells, which were incubated with 0.25 mM NiCl2 for 0, 3, 6, and 24 h. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31963541), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Angiopoietin-like Protein 4/ANGPTL4 by Western Blot NiCl2 activates the expression of ANGPTL4 and HIF1-alpha in BEAS-2B cells. (A) ANGPTL4 protein expression of BEAS-2B cells exposed to NiCl2 (0, 0.06, 0.12, and 0.25 mM) for 24 h was performed using western blotting. (B) The mRNA level of cells exposed to NiCl2 for 6 h was performed by real time-PCR. Significant differences from the untreated cells are indicated by ** p < 0.01 or *** p < 0.001 (C,D) Time-course analysis was also performed on BEAS-2B cells, which were incubated with 0.25 mM NiCl2 for 0, 3, 6, and 24 h. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31963541), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Angiopoietin-like Protein 4/ANGPTL4 by Western Blot Metformin represses NiCl2-induced ANGPTL4 activation and hypoxia-inducible factor-1 alpha (HIF-1 alpha ) expression. (A) Protein profiles were performed in NiCl2- and metformin-treated BEAS-2B cells for 48 h using a Proteome Profiler Human XL Oncology Array kit. (B) Seventeen gene expression values were generated by calculating the mean spot pixel density from the array that was affecting by NiCl2 and metformin. Data are presented as a fold change in each protein compared with the untreated group. (C,D) BEAS-2B and (E,F) A549 cells combined with 0.25 mM or 1 mM NiCl2 and 5 mM or 10 mM metformin for 24 h, western blotting and qPCR were used to determine protein and mRNA expression. (G) Migration of A549 cells across transwell filters for 24 h. A quantity of 10% FBS DMEM medium was placed in the bottom chamber containing various doses of ANGPTL4 recombinant protein. Bar graph: Quantitation of migrated cells/field. The results were obtained from three random fields per filter and represented the means ± SD. *** p < 0.001 (H) Cells were pretreated with 0, 2.5, and 5 mM metformin for 24 h and then seeded into transwell chambers without metformin; the bottom chamber contained 25 ng/mL ANGPTL4 recombinant protein. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31963541), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Angiopoietin-like Protein 4/ANGPTL4 by Western Blot Analysis of ANGPTL4 expression in lung cancer cell lines and normal cell lines exposed to NiCl2. (A) Protein expression of ANGPTL4 in various normal and tumour lung cells was performed using Western blot analysis. beta -actin was used as an internal control. (B) ANGPTL4 expression of NiCl2 derived from various cancer and normal cell lines by Western blotting and (C) real time-PCR. For protein levels, beta -actin was used as an internal control. The relative ratio of ANGPTL4 to beta -actin is shown. For mRNA levels, quantification of ANGPTL4 was normalised to beta -actin, with an average of three independent readings (mean ± SD). *** p < 0.001 versus control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31963541), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Angiopoietin-like Protein 4/ANGPTL4 by Western Blot Metformin represses NiCl2-induced ANGPTL4 activation and hypoxia-inducible factor-1 alpha (HIF-1 alpha ) expression. (A) Protein profiles were performed in NiCl2- and metformin-treated BEAS-2B cells for 48 h using a Proteome Profiler Human XL Oncology Array kit. (B) Seventeen gene expression values were generated by calculating the mean spot pixel density from the array that was affecting by NiCl2 and metformin. Data are presented as a fold change in each protein compared with the untreated group. (C,D) BEAS-2B and (E,F) A549 cells combined with 0.25 mM or 1 mM NiCl2 and 5 mM or 10 mM metformin for 24 h, western blotting and qPCR were used to determine protein and mRNA expression. (G) Migration of A549 cells across transwell filters for 24 h. A quantity of 10% FBS DMEM medium was placed in the bottom chamber containing various doses of ANGPTL4 recombinant protein. Bar graph: Quantitation of migrated cells/field. The results were obtained from three random fields per filter and represented the means ± SD. *** p < 0.001 (H) Cells were pretreated with 0, 2.5, and 5 mM metformin for 24 h and then seeded into transwell chambers without metformin; the bottom chamber contained 25 ng/mL ANGPTL4 recombinant protein. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31963541), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Angiopoietin-like Protein 4/ANGPTL4 by Western Blot Analysis of ANGPTL4 expression in lung cancer cell lines and normal cell lines exposed to NiCl2. (A) Protein expression of ANGPTL4 in various normal and tumour lung cells was performed using Western blot analysis. beta -actin was used as an internal control. (B) ANGPTL4 expression of NiCl2 derived from various cancer and normal cell lines by Western blotting and (C) real time-PCR. For protein levels, beta -actin was used as an internal control. The relative ratio of ANGPTL4 to beta -actin is shown. For mRNA levels, quantification of ANGPTL4 was normalised to beta -actin, with an average of three independent readings (mean ± SD). *** p < 0.001 versus control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31963541), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Angiopoietin-like Protein 4/ANGPTL4 by Western Blot HIF-1 alpha is involved in NiCl2-induced ANGPTL4 expression. (A) N-acetyl-L-cysteine (NAC), the ROS scavenger, inhibited NiCl2-upregulated ANGPTL4 expression. Determination of HIF-1 alpha and ANGPTL4 protein levels in BEAS-2B cells pretreated with NAC for 1 h and cultured under NiCl2 for 24 h. (B) qPCR was performed to detect ANGPTL4 mRNA levels. Data are shown relative to untreated cells after normalisation to beta -actin and reported as the mean ± SD of triplicate experiments. *** p < 0.001 compared with untreated cells; ###p < 0.001 compared with NiCl2-treated cells, as determined by two-tailed t tests. (C) Cells were pretreated with PTX-478, the HIF-1 alpha transcriptional inhibitor, for 1 h and then exposed to NiCl2 for 24 h. Western blotting and (D) qPCR were performed to determine the protein and mRNA expression. (E) BEAS-2B cells were treated with NiCl2 and metformin for 48 h after infecting with lentivirus carrying shGFP (vector control) or shHIF-1 alpha (clone no. #808 and #810). The protein levels of HIF-1 alpha, ANGPTL4, and beta -actin were determined while using Western blot analysis. beta -actin was used as the internal control. (F) Quantitative analysis of ANGPTL4 mRNA was performed using qPCR. The data are presented as the mean ± SD of triplicate experiments; a = p < 0.001 compared with shGFP untreated cells; b = p < 0.001 as compared with shGFP Ni-treated cells; c = p < 0.05 compared with shGFP Ni- and metformin cotreated cells. (G) HIF-1 alpha overexpression promoted ANGPTL4, which was then inhibited by metformin. Western blot analysis of BEAS-2B cells overexpressing pcDNA3 or HA-HIF-1 alpha -pcDNA3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31963541), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Angiopoietin-like Protein 4/ANGPTL4 by Western Blot HIF-1 alpha is involved in NiCl2-induced ANGPTL4 expression. (A) N-acetyl-L-cysteine (NAC), the ROS scavenger, inhibited NiCl2-upregulated ANGPTL4 expression. Determination of HIF-1 alpha and ANGPTL4 protein levels in BEAS-2B cells pretreated with NAC for 1 h and cultured under NiCl2 for 24 h. (B) qPCR was performed to detect ANGPTL4 mRNA levels. Data are shown relative to untreated cells after normalisation to beta -actin and reported as the mean ± SD of triplicate experiments. *** p < 0.001 compared with untreated cells; ###p < 0.001 compared with NiCl2-treated cells, as determined by two-tailed t tests. (C) Cells were pretreated with PTX-478, the HIF-1 alpha transcriptional inhibitor, for 1 h and then exposed to NiCl2 for 24 h. Western blotting and (D) qPCR were performed to determine the protein and mRNA expression. (E) BEAS-2B cells were treated with NiCl2 and metformin for 48 h after infecting with lentivirus carrying shGFP (vector control) or shHIF-1 alpha (clone no. #808 and #810). The protein levels of HIF-1 alpha, ANGPTL4, and beta -actin were determined while using Western blot analysis. beta -actin was used as the internal control. (F) Quantitative analysis of ANGPTL4 mRNA was performed using qPCR. The data are presented as the mean ± SD of triplicate experiments; a = p < 0.001 compared with shGFP untreated cells; b = p < 0.001 as compared with shGFP Ni-treated cells; c = p < 0.05 compared with shGFP Ni- and metformin cotreated cells. (G) HIF-1 alpha overexpression promoted ANGPTL4, which was then inhibited by metformin. Western blot analysis of BEAS-2B cells overexpressing pcDNA3 or HA-HIF-1 alpha -pcDNA3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31963541), licensed under a CC-BY license. Not internally tested by R&D Systems.