Cells were fixed in −20 °C cold methanol for 10 minutes then blocked with phosphate buffered saline containing 5% donkey serum at room temperature for 30 minutes. For sectioning of 3D collagen 3T3-L1, collagen plug was embedded in Optimal Cutting Temperature compound (Sakura Finetek USA, Torrance, CA) as previously described (Lee et al., 2004 (link)). Sectioning was performed at University of California–San Diego Moores Histology Core (La Jolla, CA). The detailed immunostaining method has been described (Sato et al., 2016 (link)). Anti-pCREB, anti-perilipin, anti-decorin, and Alexa Fluor 488- or 594-conjugated donkey IgG (Thermo Fisher Scientific) were used at 1μg/ml. Cover slips were mounted using ProLong Gold Antifade Reagent with DAPI (Thermo Fisher Scientific). Images were captured using a BX41 microscope (Olympus, Center Valley, PA).
Anti perilipin
Manufactured by Thermo Fisher Scientific
Anti-perilipin is a primary antibody used for the detection and quantification of perilipin proteins, which are important structural components of lipid droplets in cells. It can be used in various techniques, such as Western blotting and immunocytochemistry, to study lipid metabolism and adipocyte biology.
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Lab products found in correlation
Anti decorin, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 or 594 conjugated donkey igg, by Thermo Fisher Scientific (1 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions)
Bx41 microscope, by Olympus (1 mentions)
Optimal cutting temperature compound, by Sakura Finetek (1 mentions)
Protein ladder, by Thermo Fisher Scientific (1 mentions)
Anti c ebpα, by Thermo Fisher Scientific (1 mentions)
Ibright 1500 imaging system, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti perilipin
1
Immunostaining of 3D Collagen Cultures
2
Western Blot Analysis of Adipogenic Markers
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Procedures of Western blotting were fully described previously (3 (link)). Primary antibodies from Cell Signaling were used: anti-adiponectin (No. 2789, 1:500), anti-C/EBPα (No. 8178S, 1:1,000), anti-PDGFRα (No. 3174S, 1:1,000), anti-perilipin (No. 9349S, 1:1,000); from Thermo Fisher Scientific: anti-collagen I (PA5-95137, 1:1,000), anti-FAPB4 (No. 710189; 1:500), anti-fibronectin (PA5-29578, 1:1,000); and from Santa Cruz: anti-PPARγ (Sc-150, 1:200). Anti-rabbit (Cell Signaling, 1:4,000, No. 7074S) or anti-mouse (Cell Signaling, 1:4,000, No. 7076S) were used as secondary antibodies and protein ladder from Thermo Fisher Scientific was used (26620). Proteins were visualized by enhanced chemiluminescence (iBright 1500 Imaging System, Invitrogen) and quantified with ImageJ Software (v.1.8.0). Ponceau coloration was used as the loading control.
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