Recombinant rat IL-1β (Peprotech), PPARδ (abcam, ab23673), Aggrecan (abcam, ab3773), Collagen II (abcam, ab239007), MMP13 (abcam, ab39012), SOX9 (abcam, ab185966), Cleaved-PARP (Cell Signaling Technology, Asp214), Cleaved-caspase3 (Cell Signaling Technology, Asp175), Bcl-2 (abcam, ab194583), Bax (Cell Signaling Technology D2E11), ATG5 (abcam, ab108327), Beclin1 (abcam, ab62557), LC3 (Sigma-Aldrich, L8918), GW501516 (Selleck, S5616), GSK3787 (Selleck, S8025).
Pparδ
Manufactured by Abcam
Sourced in United States
PPARδ is a nuclear receptor protein that functions as a transcription factor. It is involved in the regulation of cellular processes such as lipid metabolism, glucose homeostasis, and cell differentiation.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 510, by Zeiss (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Aggrecan, by Abcam (1 mentions)
Mmp13, by Abcam (1 mentions)
Recombinant rat il 1β, by Thermo Fisher Scientific (1 mentions)
Collagen 2, by Abcam (1 mentions)
Gsk3787, by Selleck Chemicals (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Bcl 2, by Abcam (1 mentions)
Beclin 1, by Abcam (1 mentions)
Rabbit anti th, by Merck Group (1 mentions)
Alexa fluor 488 anti rabbit or anti mouse, by Thermo Fisher Scientific (1 mentions)
Sds polyacrylamide gel, by Bio-Rad (1 mentions)
Pvdf membrane, by Abcam (1 mentions)
M per reagent, by Thermo Fisher Scientific (1 mentions)
Pparγ, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Pparα, by Abcam (1 mentions)
4 protocols using pparδ
1
Chondrocyte Differentiation and Apoptosis
2
Immunofluorescent Staining of PPARγ and PPARδ
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Immunofluorescent staining was performed as described (Teismann et al., 2003; Sathe et al., 2012 ). Sections were washed three times for 5 min with 0.1% Triton X in 0.1 M phosphate-buffered saline (PBS), before non-specific binding was blocked with 10% normal goat serum in 0.1 M PBS-Triton X (PBS-T). Sections were incubated overnight at 4 °C in 0.1 M PBS-T with primary antibodies as follows: rabbit anti-TH (1:1000; Millipore), PPARγ (1:100, Alexis Biochemicals, San Diego, CA, USA), PPARδ (1:250, Abcam, Cambridge, UK). Following further washes, immunostaining was visualized with Alexa Fluor 488 anti-rabbit or anti-mouse (1:300; Molecular Probes, Eugene, OR, USA) or cy-3 anti-rabbit or anti-mouse (1:200; Jackson ImmunoResearch, West Grove, PA, USA) antibodies. After three final washes, sections were mounted on slides with Mowiol-DABCO. Immunostaining was visualized by confocal microscopy (LSM 510 or LSM 700, Carl Zeiss).
3
Western Blot Analysis of PPAR Isoforms
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APs were lysed and total cellular protein was isolated using M-PER reagent (Thermo Scientific Inc.). Equal amounts (15μg) of protein from each sample was loaded on 4–15% SDS-polyacrylamide gel (Bio-Rad Inc., CA), separated and transferred onto a PVDF membrane (Bio-Rad Inc., CA). Expressions were detected by incubating the PVDF membrane with specific antibodies to PPAR-α (1:1000), PPAR-δ (1:1000), Tim 4(1:500) (abcam Inc., MA) and PPAR-γ (1:1000) (Cell signaling technologies Inc.). Signals were visualized by chemiluminescent detection.
4
Western Blot Analysis of PPARδ and HDAC3
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Tissue lysate was equally loaded on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in which protein migrated in an electric field according to their molecular weight. Proteins were then transferred onto polyvinylidene fluoride (PVDF) membrane by a semi-dry method. The membrane was placed on bovine serum albumin (BSA) with phosphate buffer saline with Tween 20 (PBST) at 4 °C overnight to block non-specific proteins. Subsequently, the membrane was incubated with primary antibodies; PPARδ (1:1000) and HDAC3 (1:2000) (Abcam, USA) for 1 h at a room temperature. After that, the membrane was washed with TBST again and then incubated with horseradish peroxidase (HRP) conjugated secondary antibodies according to the manufacturer's instructions. Levels of PPARδ and HDAC3 protein expression were presented as the ratio between band densities of each protein to cyclophilin B (CPB) which was used as a loading control. For the visualization, signals of all target proteins were generated using a chemiluminescence kit and the band density values were calculated using ChemiDocTM Touch Imaging System (BioRad laboratories, CL, USA).
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