Angptl4

Manufactured by Thermo Fisher Scientific

Sourced in United States

ANGPTL4 is a protein that plays a role in the regulation of lipoprotein and glucose metabolism. It is a member of the angiopoietin-like (ANGPTL) protein family. ANGPTL4 is involved in the modulation of lipoprotein lipase activity and triglyceride metabolism.

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Lab products found in correlation

Gapdh, by Santa Cruz Biotechnology (2 mentions) Orthophosphoric acid, by Thermo Fisher Scientific (2 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (2 mentions) Sodium nitrate, by Thermo Fisher Scientific (2 mentions) Sulfanilamide, by Merck Group (2 mentions) β actin, by Thermo Fisher Scientific (2 mentions) Lps o55 b5, by Merck Group (2 mentions) Gw0742, by Merck Group (2 mentions) L 165042, by Merck Group (2 mentions) Gsk0660, by Merck Group (2 mentions) Gsk3787, by Merck Group (2 mentions) Pdk 4, by Thermo Fisher Scientific (2 mentions) Z vad fmk, by Merck Group (1 mentions) Ferrostatin 1, by Merck Group (1 mentions) Liproxstatin 1, by Merck Group (1 mentions) Tazs89a, by Addgene (1 mentions) Gp91 ds tat, by Eurogentec (1 mentions) Vinculin, by Santa Cruz Biotechnology (1 mentions) Yap taz, by Cell Signaling Technology (1 mentions) V5 tag, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Human ANGPTL4 ELISA kit (2 citations) ANGPTL4/FIAF (PA1-1053) (2 citations)

8 protocols using angptl4

Molecular Mechanisms of Ferroptosis Regulation

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Erastin was obtained from the Duke University Small Molecule Synthesis Facility. The following antibodies, their catalog numbers, sources and diltuionswere indicated below: YAP/TAZ (#8418, Cell Signaling Technology, 1:1000), βa-tubulin (#86298, Cell Signaling Technology, 1:2000), vinculin (sc-73614, Santa Cruz, 1:2000), V5 tag (#13202, Cell Signaling Technology, 1:2000), H3 (#4499, Cell Signaling Technology, 1:2000), GAPDH (sc-25778, Santa Cruz, 1:2000), ANGPTL4 (#40–9800, ThermoFisher Scientific, 1:1000), NOX2 (sc-130543, Santa Cruz, 1:1000), anti-rabbit IgG, horseradish peroxidase (HRP)-linked antibody (#7074, Cell Signaling Technology, 1:2000–1:4000) and anti-mouse IgG, HRP-linked Antibody (#7072, Cell Signaling Technology, 1:2000–1:4000). Plasmids were obtained from Addgene (TAZS89A #52084; ANGPTL4-V5 #102446). The NOX2 inhibitor, gp91 ds-tat, was purchased from Eurogentec (cat #: AS-63818) and recombinant human ANGPTL4 protein was purchased from Novus Biologicals (4487-AN). VAS2870 (Calbiochem-492000), GKT136901 (Calbiochem-534032), Z-VAD-FMK (SML0583), ferrostatin-1 (SML0583) and liproxstatin-1 (SML1414) were purchased from Sigma.

Yang W.H., Huang Z., Wu J., Ding C.K., Murphy S.K, & Chi J.T. (2019). A TAZ-ANGPTL4-NOX2 axis regulates ferroptotic cell death and chemoresistance in epithelial ovarian cancer. Molecular cancer research : MCR, 18(1), 79-90.

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Protein Expression Analysis in Pancreatic Cells

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The pancreas and pancreatic acinar cells were lysed with a radioimmunoprecipitation assay (RIPA) buffer (Biosesang, Gyeonggi‐do, Korea) containing protease and phosphatase inhibitor cocktails (GenDepot, Barker, TX). The proteins were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS‐PAGE) and transferred onto nitrocellulose membranes. The blots were immunostained with the appropriate primary antibodies followed by secondary antibodies conjugated to horseradish peroxidase. Antibody binding was detected with an enhanced chemiluminescence reagent (Bio‐Rad. Hercules, CA, USA). The primary antibodies against the following factors were used: ANGPTL4 (Thermo Fisher Scientific, Waltham, MA, USA, cat# 40‐9800), GAPDH, and β‐actin (Santa Cruz Biotechnology, Dallas, TX, USA, cat# sc‐47724, and sc‐47778). The secondary antibodies were purchased from Santa Cruz Biotechnology.

Jung K.H., Son M.K., Yan H.H., Fang Z., Kim J., Kim S.J., Park J.H., Lee J.E., Yoon Y., Seo M.S., Han B.S., Ko S., Suh Y.J., Lim J.H., Lee D., Teo Z., Wee J.W., Tan N.S, & Hong S. (2020). ANGPTL4 exacerbates pancreatitis by augmenting acinar cell injury through upregulation of C5a. EMBO Molecular Medicine, 12(8), e11222.

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Protein Expression Analysis by Western Blot

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Cells were lysed in RIPA buffer (Sefton 2001 ), and total cell proteins were separated in a 6%–15% denaturing polyacrylamide gel, transferred to nitrocellulose (Amersham Biosciences), and probed using antibodies recognizing TSN (Abcam), UPF1 (Serin et al. 2001 (link)), Flag (Sigma), MYC (Calbiochem), AGO2 (Abcam), GW182 (Bethyl Laboratories), PABPC1 (Santa Cruz Biotechnology), calnexin (Enzo Life Sciences), HIS (Qiagen), GAPDH (Santa Cruz Biotechnolgy), ITGA5 (Abcam), ITGA2 (Abcam), ANGPTL4 (Thermo Fisher Scientific), ADAM9 (Abcam), or Dicer (Abcam). Blots were quantified using Image Quant (Molecular Dynamics).

Elbarbary R.A., Miyoshi K., Hedaya O., Myers J.R, & Maquat L.E. (2017). UPF1 helicase promotes TSN-mediated miRNA decay. Genes & Development, 31(14), 1483-1493.

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Quantitative PCR Analysis of Endothelial Genes

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Total RNA was isolated from HUVEC or sorted bone ECs using RNA Plus mini kit (Qiagen, Cat#74134) according to the manufacturer’s instructions. RNA was reverse-transcribed using the iScript cDNA synthesis kit (Bio-Rad, Cat#1708890). Quantitative PCR was carried out using gene TaqMan Gene Expression Master Mix (ThermoFisher Scientific, Cat#4369016) and specific Taqman probes human: eukaryotic 18S rRNA (4319413E), Vegfa (Hs00900055_m1, Angptl4 (Hs01101127_m1), IGFBP2 (Hs01040719_m1), XBP1 (Hs00231936_m1), CTGF (Hs01026927_g1), CYR61(Hs00998500_g1), YAP1(Hs00902712_g1), WWTR1(Hs00210007_m1), HIF1A(Hs00153153_m1) and mouse probes: Vegfa (Mm00437306_m1), Angptl4 (Mm00480431_m1), Ctgf (Mm01192932_g1), Cyr61 (Mm00487499_g1) from ThermoFisher Scientific (Cat# 4331182) using a C1000 Touch Thermal cycler (BIORAD).
Primer sequences for qPCR analysis of Yap1 and Wwtr1 expression in freshly isolated bone ECs (Figure 2—figure supplement 1B) are provided in the Key Resources Table (Supplementary file 1).

Sivaraj K.K., Dharmalingam B., Mohanakrishnan V., Jeong H.W., Kato K., Schröder S., Adams S., Koh G.Y, & Adams R.H. (2020). YAP1 and TAZ negatively control bone angiogenesis by limiting hypoxia-inducible factor signaling in endothelial cells. eLife, 9, e50770.

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Western Blotting of Cell Lysates

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Western blotting using 12% SDS‒PAGE was performed, and 30 μg of cell lysates were analyzed as previously described 12 (link). Antibodies against human NOX4 (GTX121929) and pCHOP Ser30 (GTX55401) (GeneTex, Hsinchu, TWN); ANGPTL4 (#409800) and pFAK Tyr397 (44-624G) (Invitrogen) (BD Transduction Laboratory, Los Angeles, CA, USA); p41/42 MAPK (#9102), pAKT Ser473 (4051S) and pEGFR Y1068 (3777S) (Cell Signaling Technology, Inc., Danvers, MA, USA); GAPDH (sc-32233) and EGFR (A-10) (sc-373746) (Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA); CXCL8 (MAB208) (R&D); CHOP (A20987) (Abclonal); and NOX4 (ab109225) (Abcam) were used as the primary antibodies.

Shen C.J., Chan R.H., Lin B.W., Li N.C., Huang Y.H., Chang W.C, & Chen B.K. (2023). Oleic acid-induced metastasis of KRAS/p53-mutant colorectal cancer relies on concurrent KRAS activation and IL-8 expression bypassing EGFR activation. Theranostics, 13(13), 4650-4666.

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Transient siRNA and Plasmid Transfection

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Transient transfection of cells with 20 nM siRNA oligonucleotides or plasmids was performed using RNAiMAX or Lipofectamine 2000 (Invitrogen) according to the manufacturer's instruction with slight modifications. The siRNA IDs were as follows, ANGPTL4 (siRNA IDs: HSS181878, HSS181879); NOX4 (siRNA IDs: HSS121312, HSS121313); MMP-1 (siRNA IDs: HSS106609, HSS106610); MMP-3 (siRNA IDs: S8854, S8855); MMP-9 (siRNA IDs: S8862, S8863); c-Jun (siRNA IDs: HSS105641) (Invitrogen); Negative control siRNAs (siRNA IDs: D-001810-10-50) (Dharmacon, Lafayette, CO, USA). For use in transfection, 3.75 μl of RNAiMAX or Lipofectamine 2000 was incubated with siRNA or plasmid in 1.5 ml of Opti-MEM medium (Invitrogen) for 30 min at room temperature. Following the removal of Opti-MEM medium and replacement with 3 ml of fresh culture medium, cells were incubated for an additional 24 h, unless stated otherwise.

Shen C.J., Chang K.Y., Lin B.W., Lin W.T., Su C.M., Tsai J.P., Liao Y.H., Hung L.Y., Chang W.C, & Chen B.K. (2020). Oleic acid-induced NOX4 is dependent on ANGPTL4 expression to promote human colorectal cancer metastasis. Theranostics, 10(16), 7083-7099.

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Ligand-Induced PPARβ/δ Signaling

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The PPARβ/δ ligands GW0742, GSK3787, L−165042 and GSK0660 as well as 1400W, LPS O55:B5, sulfanilamide and naphthylethylenediamine dihydrochloride were purchased from Sigma (Gillingham, Dorset, UK). Sodium nitrate, orthophosphoric acid and DMSO were purchased from Fisher Scientific (Loughborough, UK). Primers from Applied Biosystem (Foster City, CA, USA): β-actin (Rn00667869_m1), Pdk−4 (Rn00585577_m1), Angptl−4 (Rn015228817_m1), Nos2 (Rn00561646_m1).

Perez Diaz N., Lione L.A., Hutter V, & Mackenzie L.S. (2021). Co-Incubation with PPARβ/δ Agonists and Antagonists Modeled Using Computational Chemistry: Effect on LPS Induced Inflammatory Markers in Pulmonary Artery. International Journal of Molecular Sciences, 22(6), 3158.

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Ligand-Activated PPAR Signaling Pathway

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The PPARβ/δ ligands GW0742, GSK3787, L-165042 and GSK0660 as well as 1400W, LPS O55:B5, sulfanilamide and naphthylethylenediamine dihydrochloride were purchased from Sigma. Sodium nitrate, orthophosphoric acid and DMSO were purchased from Fisher Scientific.
Primers from Applied Biosystem: β-actin (Rn00667869_m1), Pdk-4 (Rn00585577_m1), Angptl-4 (Rn015228817_m1), Nos2 (Rn00561646_m1), Serpine-1 (Rn01481341_m1), Timp-1 (Rn01430873_m1), Id2 (Rn01495280_m1).

Diaz N.P., Lione L.A., Hutter V., & Mackenzie L.S. (2020). Modelling the effects of PPARβδ of innate inflammatory responses in lung tissues.

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