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Hyperfilm ecl film

Manufactured by Euroclone
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Sourced in Italy

Hyperfilm-ECL film is a chemiluminescent detection film used for capturing and visualizing signals in Western blot and other immunoassay techniques. It is designed to be compatible with enhanced chemiluminescence (ECL) detection systems.

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Lab products found in correlation

Liteablot turbo luminol reagents, by Euroclone (5 mentions) Mouse cytokine array panel a kit, by R&D Systems (3 mentions) Cell lysis buffer, by Cell Signaling Technology (2 mentions) Rabbit anti cxcr4, by Abcam (1 mentions) Rabbit anti traf6, by Abcam (1 mentions) Rabbit anti osterix, by Santa Cruz Biotechnology (1 mentions) Rabbit anti tgf β, by Abcam (1 mentions) Rabbit anti cxcl12, by Abcam (1 mentions) Mouse anti rankl, by Abcam (1 mentions) Hrp conjugated rabbit anti mouse igg, by Cell Signaling Technology (1 mentions) Anti α tubulin, by Merck Group (1 mentions)

5 protocols using hyperfilm ecl film

1

Cytokine Profile Analysis in BMCs

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The cytokine/chemokine profiles in supernatants of 3-day cultured BMCs population were assessed by using Mouse Cytokine Array Panel A kit (R&D Systems, Milano, Italy) accordingly to the manufacturer’s instructions. Briefly, after 3 days in culture BMCs supernatant (600 μl) from each experimental group was incubated overnight on nitrocellulose membranes spotted with specific antibodies. Chemiluminescence detection produced signals (blots) directly proportional to the amount of cytokine bound. Immunoreactive dots were visualized using LiteAblot Turbo luminol reagents (EuroClone, Milano, Italy) and Hyperfilm-ECL film (EuroClone, Milano, Italy) and quantitated densitometrically using Image J software.
Sabbieti M.G., Lacava G., Amaroli A., Marchetti L., Censi R., Di Martino P, & Agas D. (2017). Molecular Adjuvants Based on Plasmids Encoding Protein Aggregation Domains Affect Bone Marrow Niche Homeostasis. Current Gene Therapy, 17(5), 391-397.
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2

Cytokine/Chemokine Profiling of BMCs

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The cytokine/chemokine profiles of BMCs supernatants were assessed by ELISA-based cytokine array by using Mouse Cytokine Array Panel A kit (R&D Systems, Milano, Italy) accordingly to the manufacturer's instructions. Immunoreactive dots were visualized using LiteAblot Turbo luminol reagents (Euroclone, Milano, Italy) and Hyperfilm-ECL film (Euroclone, Milano, Italy) and quantitated densitometrically.
Sabbieti M.G., Agas D., Capitani M., Marchetti L., Concetti A., Vullo C., Catone G., Gabai V., Shifrin V., Sherman M.Y., Shneider A, & Venanzi F.M. (2015). Plasmid DNA-coding p62 as a bone effective anti-inflammatory/anabolic agent. Oncotarget, 6(6), 3590-3599.
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3

Quantifying CXCL12 and CXCR4 Protein Levels

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Proteins from BMCs were extracted in cell lysis buffer (Cell Signaling, EuroClone, Milano, Italy) after 3 days of culture, and the concentration was determined by the BCA protein assay reagent (Pierce, EuroClone). Western blotting was performed as previously described [14 (link)]. Membranes were immunoblotted in blocking buffer with rabbit anti-CXCL12 or with rabbit anti-CXCR4 (Abcam, Prodotti Gianni, Milano, Italy) both diluted 1:600. After washing with PBS-T, blots were incubated with horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG IgG (Cell Signaling, EuroClone) diluted 1:100,000.
Immunoreactive bands were visualized using LiteAblot Turbo luminol reagents (EuroClone) and Hyperfilm- ECL film (EuroClone) according to the manufacturer’s instructions. To normalize the bands, filters were stripped and re-probed with a monoclonal anti-α-tubulin (Sigma-Aldrich). Band density was quantified densitometrically.
Sabbieti M.G., Lacava G., Amaroli A., Marchetti L., Censi R., Di Martino P, & Agas D. (2017). Molecular Adjuvants Based on Plasmids Encoding Protein Aggregation Domains Affect Bone Marrow Niche Homeostasis. Current Gene Therapy, 17(5), 391-397.
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4

Cytokine and Chemokine Profiling of BMCs

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The cytokine/chemokine profiles in supernatants of 2-day cultured BMCs population as well as in serum samples were assessed by using Mouse Cytokine Array Panel A kit (R&D Systems) according to the manufacturer's instructions. Immunoreactive dots were visualized using LiteAblot Turbo luminol reagents (EuroClone, Milano, Italy) and Hyperfilm-ECL film (EuroClone) and quantitated densitometrically.
Agas D., Gusmão Silva G., Laus F., Marchegiani A., Capitani M., Vullo C., Catone G., Lacava G., Concetti A., Marchetti L, & Sabbieti M.G. (2017). INF-γ encoding plasmid administration triggers bone loss and disrupts bone marrow microenvironment. The Journal of endocrinology, 232(2).
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5

Western Blot Analysis of Bone Cells

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Proteins from total BMCs and from BMSCs were extracted in cell lysis buffer (Cell Signaling, EuroClone) after 2 days of culture, and the concentration was determined by the BCA protein assay reagent (Pierce, EuroClone). Western blotting was performed as previously described (Sabbieti et al. 2010) (link). Membranes were immunoblotted in blocking buffer with specific antibodies: rabbit anti-TNFα and rabbit anti-NF-κB (BioLegend, Microtech SrL, Napoli, Italy) both diluted 1:500; mouse anti-RANKL, rabbit anti-TRAF6, rabbit anti-CXCL12 and rabbit anti-TGFβ (Abcam, Prodotti Gianni) all diluted 1:600; rabbit anti-PPRγ (Santa Cruz Biotechnology, Inc. DBA) diluted 1:300 and rabbit anti-Osterix (Santa Cruz Biotechnology, DBA) diluted 1:300. After washing with PBS-T, blots were incubated with horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG or with HRP-conjugated rabbit anti-mouse IgG (Cell Signaling, EuroClone) both diluted 1:50,000. Immunoreactive bands were visualized using LiteAblot Turbo luminol reagents (EuroClone) and Hyperfilm-ECL film (EuroClone) according to the manufacturer's instructions. To normalize the bands, filters were stripped and re-probed with a monoclonal anti-α-tubulin (Sigma-Aldrich). Band density was quantified densitometrically.
Agas D., Gusmão Silva G., Laus F., Marchegiani A., Capitani M., Vullo C., Catone G., Lacava G., Concetti A., Marchetti L, & Sabbieti M.G. (2017). INF-γ encoding plasmid administration triggers bone loss and disrupts bone marrow microenvironment. The Journal of endocrinology, 232(2).
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