A previously described procedure was used24 with the following primary antibody (SERT: 1:1000, sc‐1458, Santa Cruz Biotechnology; Flotillin‐1:1:1000, ab133497, Abcam; β‐actin: 1:2000, A0760‐40, US Biological, Salem, Massachusetts) which were incubated with membranes overnight at 4°C. Target protein densitometry values were normalized to those of the housekeeping protein to semi‐quantitatively determine protein levels.
Flotillin
Manufactured by Abcam
Sourced in United States
Flotillin is a membrane-associated protein that is involved in the formation and organization of lipid rafts in the cell membrane. It is commonly used as a marker for lipid rafts in biochemical and cell biology research.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse, by Santa Cruz Biotechnology (2 mentions)
Calnexin, by Abcam (2 mentions)
Glial fibrillary acidic protein (gfap), by Merck Group (2 mentions)
Goat anti rabbit, by Santa Cruz Biotechnology (2 mentions)
β actin, by Merck Group (2 mentions)
Ab133497, by Abcam (1 mentions)
β actin, by US Biological (1 mentions)
Sc 1458, by Santa Cruz Biotechnology (1 mentions)
A0760 40, by US Biological (1 mentions)
Ab201060, by Abcam (1 mentions)
Ab10148, by Abcam (1 mentions)
Alexa fluor 594 conjugated goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 647 conjugated goat anti mouse, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 conjugated goat anti mouse, by Thermo Fisher Scientific (1 mentions)
Synaptophysin, by Abcam (1 mentions)
Gephyrin, by Abcam (1 mentions)
Gad65, by Abcam (1 mentions)
Vglut1, by Merck Group (1 mentions)
Drebrin, by Abcam (1 mentions)
Hif 1α sirna, by Santa Cruz Biotechnology (1 mentions)
5 protocols using flotillin
1
Protein Expression Quantification Protocol
2
Immunochemical Analysis of Alzheimer's Markers
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Following antibodies were used for this study. AβmOC64 (ab201060, Abcam, MA, USA), Aβ 1-42 (ab10148,), BACE1 (ab63954, Abcam, MA, USA), APP (ab15272, Abcam, MA, USA) CD63 (ab216130, Abcam, MA, USA), CD9 (ab92726, Abcam, MA, USA), Flotillin (ab133497, Abcam, MA, USA), TSG101 (ab125011, Abcam, MA, USA), Alix (ab275377, Abcam, MA, USA), Calnexin (ab133615, Abcam, MA, USA), Argonuate-2 (ab186733, Abcam, MA, USA), GFAP (G3893, Sigma- Aldrich, MO, USA), β-actin (A5316, Sigma- Aldrich, MO, USA), HIF-1α (NB100-449, Novus Biological Company, CO, USA), goat anti-rabbit (sc-2004, Santa Cruz Biotechnology, TX, USA) and goat anti-mouse (sc-2005, Santa Cruz Biotechnology, TX, USA), Alexa Fluor 488 conjugated goat anti-mouse (A11001, ThermoFisher Scientific, MA, USA), Alexa Fluor 647 conjugated goat anti-mouse (A32728, ThermoFisher Scientific, MA, USA), Alexa Fluor 594 conjugated goat anti-rabbit (A11012, ThermoFisher Scientific, MA, USA).
3
Molecular Markers for Neurodegenerative Diseases
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The following reagents and antibodies were used for this study: HIV‐1 Tat 101 (Immunodiaognistics, Woburn, MA), mutant Tat (ab83352, Abcam, MA, USA), HIF‐1α siRNA (sc‐35561, Santa Cruz Biotechnology, TX, USA), GW4869 (D1692, Sigma‐Aldrich, MO, USA), GFP‐plasmid (13031, Adgene, MA, USA), antibodies‐ AβmOC64 (ab201060, Abcam, MA, USA), APP (ab15272, Abcam, MA, USA), TSG101 (ab125011, Abcam, MA, USA), Ago‐2 (ab186733, Abcam, MA, USA), Alix (ab275377, Abcam, MA, USA), Calnexin (ab133615, Abcam, MA, USA), Flotillin (ab133497, Abcam, MA, USA), CD63 (ab216130, Abcam, MA, USA), Drebrin (ab178408, Abcam, MA, USA), GAD65 (ab239372, Abcam, MA, USA), Gephyrin (ab181382, Abcam, MA, USA), GFAP (G3893, Sigma‐Aldrich, MO, USA), Goat anti‐mouse (sc‐2005, Santa Cruz Biotechnology, TX, USA), Goat anti‐rabbit (sc‐2004, Santa Cruz Biotechnology, TX, USA), HIF‐1α (NB100‐449, Novus Biological Company, CO, USA), Iba1 (ab17884, Abcam, MA, USA) IL‐6 (ab179570, Abcam, MA, USA), Tau (ab9778, Abcam, MA, USA), p‐tau 214 (ab170892, Abcam, MA, USA), p‐Tau 622 (ab76128, Abcam, MA, USA), Synaptophysin (ab8049 Abcam, MA, USA), vGLUT1 (AB5905, Millipore, MA, USA), NeuN (ab134014, Abcam, MA, USA), MAP 2 (ab5392, Abcam, MA, USA), β‐Actin (A5316, Sigma‐Aldrich, MO, USA) were purchased from commercial vendors.
4
Immunoblotting of Apoptosis Regulators
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S-100 cytosol and pellet fractions or lipid raft fractions were denatured in loading buffer and resolved on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by transfer to PVDF membranes. Immunoblotting was performed with antibodies against Apaf-1 (Chemicon, CA), β-actin, Bcl-2, Bcl-xl, VDAC-1 (Santa-Cruz, Dallas, TX), BCL11A, caveolin and flotillin (Abcam, Cambridge, UK) and then probed with the isotype specific secondary IgG-HRP as required. Membranes were then exposed to Super Signal Substrate Western Blotting Kit (Pierce, Rockford, IL, USA).
5
Western Blot Analysis of Exosomal Proteins
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Heart tissues, exosomes, and cells were lysed using RIPA buffer (Beyotime, Jiangsu). Protein concentration was measured by the BCA method (Thermo Fisher Scientific, USA). Equal amounts of protein were loaded on SDS-PAGE gels, separated, and then transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA). Then, the membranes were incubated with primary rabbit anti-mouse antibodies (CD9, flotillin, Hsp70; YAP1, bcl-2, cleaved-caspase 3, CTGF, TEAD1, ANP, and β-actin; Abcam; USA) at a dilution of 1:1,000. After overnight incubation at 4°C, the membranes were subsequently incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG; 1:10,000) for 1 h at room temperature. The immunobands were visualized using an enhanced chemiluminescence (ECL) detection kit (Beyotime, PR China).
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