Perilipin

Manufactured by Merck Group

Sourced in France

Perilipin is a lab equipment product manufactured by Merck Group. It is a protein that plays a core function in the regulation of lipid storage and metabolism in cells.

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Lab products found in correlation

6 protocols using perilipin

Tissue Preparation and Immunohistochemistry of Mouse Limbs

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Full-thickness skin was dissected from mouse upper forelimb or lower back and attached to filter paper before submersing in 4% PFA in phosphate buffer. Tissue was fixed overnight at 4°C and then washed in PBS and serial overnight washes in 10% and 30% sucrose in PBS before being embedded in OCT and cryo-sectioned in longitudinal sections of 12 µm. For mouse limbs from E18.5–52 weeks of age, limbs were harvested and fixed overnight in PFA, washed in PBS, and incubated with 400 µM EDTA in PBS for approximately 1 week before washing in sucrose, embedding and cryo-sectioning. Standard immunohistochemistry techniques were used for staining. Primary antibodies: PRRX1 (MPI-CBG Antibody facility) used at 1:200, SOX9 (R&D Systems, AF3075) used at 1:200, PERILIPIN (Sigma-Aldrich, P1873) used at 1:200, CD31 (BD Pharmingen, 557355) used at 1:100.

Currie J.D., Grosser L., Murawala P., Schuez M., Michel M., Tanaka E.M, & Sandoval-Guzmán T. (2019). The Prrx1 limb enhancer marks an adult subpopulation of injury-responsive dermal fibroblasts. Biology Open, 8(7), bio043711.

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Western Blot Analysis of Myogenic Regulators

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Western blot was performed using antibodies against the following proteins: BAF60A (BD), BAF60B (Santa Cruz Biotechnology), BAF60C (Forcales et al. 2012 (link)), MYOD (Santa Cruz Biotechnology), MYHC (MF20, Developmental Studies Hybridoma Bank [DSHB]), PERILIPIN (Sigma-Aldrich), and TUBULIN (Cell Signaling). Further experimental details are provided in the Supplemental Material.

Saccone V., Consalvi S., Giordani L., Mozzetta C., Barozzi I., Sandoná M., Ryan T., Rojas-Muñoz A., Madaro L., Fasanaro P., Borsellino G., De Bardi M., Frigè G., Termanini A., Sun X., Rossant J., Bruneau B.G., Mercola M., Minucci S, & Puri P.L. (2014). HDAC-regulated myomiRs control BAF60 variant exchange and direct the functional phenotype of fibro-adipogenic progenitors in dystrophic muscles. Genes & Development, 28(8), 841-857.

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Immunohistochemical Analysis of Bone Tissue

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Harvested femurs were decalcified by 0.5 M EDTA. The tissues were embedded in gelatin (at a concentration of 8%) with sucrose (at a concentration of 20%) in the presence of polyvinylpyrrolidone (at a concentration of 2%). We sliced the tissues into 20 μm-thick sections. Subsequently, we blocked the samples, treated the samples with primary antibodies (the antibodoy for TAZ, 1:200, Abcam; for OCN, 1:200, R&D; for perilipin, 1:400, Sigma), and incubated them with secondary antibodies.

Wang N., Li Z., Li S., Li Y., Gao L., Bao X., Wang K., Liu C., Xue P, & Liu S. (2021). Curculigoside Ameliorates Bone Loss by Influencing Mesenchymal Stem Cell Fate in Aging Mice. Frontiers in Cell and Developmental Biology, 9, 767006.

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Comprehensive Protein Analysis of Adipogenesis

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SDS-PAGE and immunoblot analysis of samples were performed using the following antibodies: custom-made PDGF D (22 (link)), fatty acid synthase (Cell Signaling Technologies [CST]; catalog no.: 3180), C/EBPα (CST; catalog no.: 8178), peroxisome proliferator–activated receptor gamma (CST; catalog no.: 2435), perilipin (CST; catalog no.: 9349), FABP4 (CST; catalog no.: 2120), acetyl CoA carboxylase (CST; catalog no.: 3676), β-actin (CST; catalog no.: 4970), p-AKT (CST; catalog no.: 9275), AKT (CST; catalog no.: 9272), p-JNK (CST; catalog no.: 4668), JNK (CST; catalog no.: 9258), p-ERK1/2 (CST; catalog no.: 9101), ERK1/2 (CST; catalog no.: 9102), p-β-PDGFR Y751 (CST; catalog no.: 3161), p-β-PDGFR Y771 (CST; catalog no.: 3173), β-PDGFR (CST; catalog no.: 3169), vinculin (Sigma; catalog no.: V9131), ubiquitin (CST; catalog no.: 3936), ubiquitin-K48 (Abcam; catalog no.: ab140601), ubiquitin-K63 (Abcam; catalog no.: ab179434), HUWE1 (Abcam; catalog no.: ab70161), and streptavidin–horseradish peroxidase (Abcam; catalog no.: ab7403).

Pham T., Najy A.J, & Kim H.R. (2022). E3 ligase HUWE1 promotes PDGF D-mediated osteoblastic differentiation of mesenchymal stem cells by effecting polyubiquitination of β-PDGFR. The Journal of Biological Chemistry, 298(6), 101981.

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Multipotent Progenitor Cell Differentiation

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Testes were collected from adult C57BL/6 or Tcf21^mCrem:R26R^tdTom mice and dissociated into a single-cell suspension and sorted for SCA1⁺/cKITit⁻ or SCA1⁺/TCF21^lin/cKITit⁻ cells, respectively. For adipogenic differentiation, 3 × 10⁴ cells were plated in a monolayer, cultured for 10 days (Stem ProAdipogenic differentiation kit) and stained with either Oil Red O or Perilipin (1:250, Sigma, Cat#P1873). For chondrogenic differentiation, 5 × 10⁴ cells were plated in micromass, cultured for 21 days (Stem ProChondrogenic differentiation kit) and stained with either Alcian blue or SOX9 (1:250, EMD Millipore, Cat#ABE571). For osteogenic differentiation, 1 × 10⁴ cells were plated in micromass, cultured for 14 days (StemProOsteogenicdifferentiationkit), and stained with either Alizarin red or Osterix (1:250, Abcam, Cat#ab22552)^{96 (link)}.

Shen Y.C., Shami A.N., Moritz L., Larose H., Manske G.L., Ma Q., Zheng X., Sukhwani M., Czerwinski M., Sultan C., Chen H., Gurczynski S.J., Spence J.R., Orwig K.E., Tallquist M., Li J.Z, & Hammoud S.S. (2021). TCF21+ mesenchymal cells contribute to testis somatic cell development, homeostasis, and regeneration in mice. Nature Communications, 12, 3876.

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Antibody Characterization for Invadolysin

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Antibodies used in this work were 3645/6 = INV P1-Ab (Cobbe et al., 2009) , INV C-Ab1 (Cobbe et al., 2009) , 4624 (INV.v1 downstream of signal sequence: SLGSSPPCRHHVPS), 4888 (INV.v2 N-terminal region: SGLLGLRPGPEPVAL), perilipin (Sigma-Aldrich), TIP-47 (Progen), Akt, phospho-Akt 505, phospho-p70 S6 kinase Thr 398, and phospho-4E-BP Thr37/46 (all Cell Signaling, Genosphere, Paris, France). The invadolysin peptide antibodies were generated by Genosphere. Lsd2 antibodies were generously provided by Dr. Ronald Kühnlein (Max Planck Institute, Munich) and Dr. Luis Teixeira (University of Cambridge). PKC inhibitors Ro31-8220, Cycloheximide, 3-isobutyl-l-methylxantin (IBMX), dexamethasone (DEX) and insulin were purchased from Sigma-Aldrich. PI-3 kinase (PI3K) was obtained from Cell Signaling.

Chang C.W., Abhinav K., Di Cara F., Panagakou I., Vass S, & Heck M.M. (2017). A role for the metalloprotease invadolysin in insulin signaling and adipogenesis. Biological chemistry, 398(3).

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