The regions of ischemia and penumbra in the right side of each MCAO brain (n=8 per group) were isolated on ice and homogenized in lysis buffer. Protein concentrations were measured with a Bio-Rad protein assay kit (Bio-Rad Laboratories, Hercules, CA, USA). Equivalent amounts of protein were separated on 10% SDS-PAGE gels, and transferred to polyvinylidene difluoride (PVDF) membranes. The membranes were probed with rabbit anti-PPARγ (1:1000 Cell Signaling Technology, Danvers, MA, USA) at 4°C overnight and subsequently incubated with anti-rabbit peroxidase-conjugated secondary antibody (1:2000, Sigma-Aldrich) for 1 h at room temperature. The blots were quantified and normalized with glyceraldehyde-3-phosphate dehydrogenase (GADPH) by scanning with a ScanJet scanner (Hewlett Packard, Inc. Palo Alto, CA, USA).
Scanjet scanner
Manufactured by Hewlett-Packard
Sourced in United States
The Hewlett-Packard ScanJet is a flatbed scanner that captures digital images from physical documents, photographs, and other materials. It features a high-resolution sensor and can produce scanned images of superior quality.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti pparγ, by Cell Signaling Technology (1 mentions)
Anti rabbit peroxidase conjugated secondary antibody, by Merck Group (1 mentions)
Protein assay kit, by Bio-Rad (1 mentions)
Anti mouse, by Merck Group (1 mentions)
Pvdf filter, by Merck Group (1 mentions)
Peroxide conjugated anti rabbit, by GE Healthcare (1 mentions)
Ecl system, by GE Healthcare (1 mentions)
2 protocols using scanjet scanner
1
Quantifying PPAR-gamma Expression in MCAO
2
Immunoblotting Protein Detection Protocol
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To detect p-(Tyr1150/1151)-IR, IR, GSK3, p-GSK3, GS, p-GS, PEPCK, G-6-Pase, SGLT-2 and GLUT-2, equal amounts of protein were separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) filters (Millipore, Madrid, Spain).
Membranes were probed with the corresponding primary antibody followed by incubation with peroxide-conjugated anti-rabbit (GE Healthcare, Madrid, Spain) or antimouse (Sigma, Madrid, Spain) immunoglobulin. Blots were developed with the ECL system (GE Healthcare, Madrid, Spain). Normalization of Western blot was ensured by β-actin and band quantification was carried out with a Scanjet scanner (Hewlett Packard) and the Scion Image software.
Membranes were probed with the corresponding primary antibody followed by incubation with peroxide-conjugated anti-rabbit (GE Healthcare, Madrid, Spain) or antimouse (Sigma, Madrid, Spain) immunoglobulin. Blots were developed with the ECL system (GE Healthcare, Madrid, Spain). Normalization of Western blot was ensured by β-actin and band quantification was carried out with a Scanjet scanner (Hewlett Packard) and the Scion Image software.
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