Tissues were fixed in 10% formalin, subjected to paraffin-embedded sectioning and stained with hematoxylin or immunostained with anti-UCP1 (Sigma) and anti-perilipin A (Abcam) antibodies. TUNEL (terminal deoxynucleotidyl transferase dUTP-mediated nick end labelling) (Promega) and cleaved caspase-3 (Cell signaling) immunostaining were performed per manufacturer’s instructions. Additional information is described in the Supplementary Experimental Procedures .
Anti perilipin a
Manufactured by Abcam
Sourced in Canada
Anti-perilipin A is a primary antibody that recognizes perilipin A, a lipid droplet-associated protein. Perilipin A is involved in the regulation of lipolysis and the storage of neutral lipids within adipocytes. This antibody can be used for western blotting, immunohistochemistry, and immunocytochemistry applications.
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Lab products found in correlation
4 protocols using anti perilipin a
1
Tissue Fixation and Immunostaining for UCP1 and Perilipin A
2
In Situ Hybridization and Immunofluorescence Analysis
3
In Situ Hybridization and Immunofluorescence Analysis
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Tissues were collected and fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS), dehydrated, embedded in paraffin, and sectioned at 5 to 6 μm. Whole mount samples were fixed and dehydrated according to the standard protocol. Tissues were then cut into thin strips.
For in situ hybridization, tissues were hybridized with digoxigenin-labeled probes. Signals were detected using an anti-digoxigenin antibody coupled to alkaline phosphatase. Sfrp4 (nt 183-755), Follistatin (nt 409-1075), and Fgf7 (nt 486-1088) probes were obtained as follows: Wnt5a (nt 74-1156) (Dr. Sharpe, King's College London), Dkk1 (nt 112-942) (Dr. Millar, University of Pennsylvania), Lef-1 (1176-1905) (Dr. Thesleff, University of Helsinki).
Immunofluorescence staining was performed with anti-follistatin (mouse, 1:50, Abcam) and anti-perilipin A (rabbit, 1:100, Abcam) antibodies followed by secondary antibodies conjugated to Alexa-488 and Alexa-568.
For in situ hybridization, tissues were hybridized with digoxigenin-labeled probes. Signals were detected using an anti-digoxigenin antibody coupled to alkaline phosphatase. Sfrp4 (nt 183-755), Follistatin (nt 409-1075), and Fgf7 (nt 486-1088) probes were obtained as follows: Wnt5a (nt 74-1156) (Dr. Sharpe, King's College London), Dkk1 (nt 112-942) (Dr. Millar, University of Pennsylvania), Lef-1 (1176-1905) (Dr. Thesleff, University of Helsinki).
Immunofluorescence staining was performed with anti-follistatin (mouse, 1:50, Abcam) and anti-perilipin A (rabbit, 1:100, Abcam) antibodies followed by secondary antibodies conjugated to Alexa-488 and Alexa-568.
4
Immunohistochemical Analysis of Adipose Tissue
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After fixation, AT was embedded in paraffin as described previously (22 (link), 29 (link)). At 4°C sections were incubated overnight with primary antibodies anti-Mac-2 (1:1000; Cedarlane; Burlington, ON, Canada; CL8942AP) and anti-PerilipinA (1:200; Abcam; Cambridge, UK; ab3526). Next, fluorochrome-conjugated secondary antibodies were applied for 1 h at room temperature (1:200; Invitrogen; Waltham, MA, USA). For nuclear staining DAPI (1:10,000; Thermo Fischer Scientific; Waltham, MA, USA; 62248) was used. For control stainings, same routines without primary antibodies were applied. A confocal Leica SPE microscope (Leica; Wetzlar, Germany) was used for image acquisition. Quantification of adipocyte size, interstitial macrophages and CLS density were performed semi-automatically with cellSens Software (Olympus; Hamburg, Germany) as described previously (30 (link)). H&E stainings were performed following standard routines (29 (link)).
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