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3 protocols using calnexin 2679

1

Detailed Lipid Droplet Imaging Protocol

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CIDEC-GFP was generously provided by professor Peng Li (Fudan University, China), BFP-Sec61β was a gift from Gia Voeltz (Addgene plasmid #49,154). Acipimox (A7856), Triascin C (T4540) and antibodies towards CIDEC (HPA020553) were from Sigma. LAMP1 (ab25630), and EHD2 (154,784) from abcam, perilipin-1 pS522 from Vala Sciences, LC3B (2775S), hormone-sensitive lipase (HSL) pS563 (4139S), Caveolin-1 (3267), Rab5 (46,449), Rab11 (5589), and Calnexin (2679) from Cell Signalling Technology. Alexa fluor goat α-rabbit 568(A11036), 647(A21245), α-mouse 568(A11031), 647(A21236) secondary antibodies, Bodipy 493/503 (D3922), Bodipy 558/568 C12 (D3835), Bodipy 665/676 (B3932), and Lysotracker Deep Red (L12492) were from Invitrogen. MitoTracker RED CMXRos (M7512) and Mitotracker Deep Red (M22426) were from ThermoFisher, 10 nm gold secondary antibodies from Electron Microscopy Sciences.
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2

Cellular Stress Signaling Pathway Antibodies

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Ubiquitin (sc-8017), LAMP1 (sc-20011), phospho-PERK (sc-32577), PERK (sc-13073), phospho-c-Jun (sc-822), c-Jun (sc-376488), SERCA2 (sc-376235, sc-8095), ATF6 (sc-22799), eIF2α (sc-11386), CaMKII (sc-5306), α1C/Cav1.2 (sc-16229-R), α1D/Cav1.3 (sc-25687), LC3 (sc-271625) and p62 (sc-28359) antibodies were obtained from Santa Cruz Biotechnology, Inc. IRE1α (3294), phospho-CaMKII (12716), phospho-eIF2α (3398), PERK (5683), LC3 (2775), p62 (5114) and Calnexin (2679) antibodies were purchased from Cell Signaling Technology, Inc. Pan-keratin antibody was obtained from Dr. Omary (University of Michigan)65 (link). Actin (JLA20) and tubulin (T5168) antibodies were purchased from Developmental Studies Hybridoma Bank (DSHB) and Sigma, respectively. Fatty acid free and low endotoxin bovine serum albumin (BSA), palmitic acid (PA), stearic acid (SA), oleic acid (OA), docosahexaenoic acid (DHA), tunicamycin (Tm), thapsigargin (Tg), butylated hydroxyanisole (BHA), N-acetylcysteine (NAC), chlorocresol, L-cycloserine, glucagon, phenylephrine, vasopressin, verapamil and nicardipine were purchased from Sigma. SP600125, bafilomycin and rapamycin were from LC Labs, and fumonisin B1 and suramin were from Cayman Chemical.
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3

Cellular Stress Signaling Pathway Antibodies

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Ubiquitin (sc-8017), LAMP1 (sc-20011), phospho-PERK (sc-32577), PERK (sc-13073), phospho-c-Jun (sc-822), c-Jun (sc-376488), SERCA2 (sc-376235, sc-8095), ATF6 (sc-22799), eIF2α (sc-11386), CaMKII (sc-5306), α1C/Cav1.2 (sc-16229-R), α1D/Cav1.3 (sc-25687), LC3 (sc-271625) and p62 (sc-28359) antibodies were obtained from Santa Cruz Biotechnology, Inc. IRE1α (3294), phospho-CaMKII (12716), phospho-eIF2α (3398), PERK (5683), LC3 (2775), p62 (5114) and Calnexin (2679) antibodies were purchased from Cell Signaling Technology, Inc. Pan-keratin antibody was obtained from Dr. Omary (University of Michigan)65 (link). Actin (JLA20) and tubulin (T5168) antibodies were purchased from Developmental Studies Hybridoma Bank (DSHB) and Sigma, respectively. Fatty acid free and low endotoxin bovine serum albumin (BSA), palmitic acid (PA), stearic acid (SA), oleic acid (OA), docosahexaenoic acid (DHA), tunicamycin (Tm), thapsigargin (Tg), butylated hydroxyanisole (BHA), N-acetylcysteine (NAC), chlorocresol, L-cycloserine, glucagon, phenylephrine, vasopressin, verapamil and nicardipine were purchased from Sigma. SP600125, bafilomycin and rapamycin were from LC Labs, and fumonisin B1 and suramin were from Cayman Chemical.
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