Cultured cells or tissues were prepared with the T-PER tissue protein extraction reagent (2% SDS and 60 mM Tris HCl, pH 6.8) with the cocktail of proteinase inhibitors (Roche, Indianapolis, IN, USA) in it. The total protein tissue or cultured cells were homogenized in lysis buffer and were loaded onto the gel (20–40 μg) for electrophoresis. Proteins then were transferred onto 10% SDS-PAGE (sodium dodecylsulfate polyacrylamide gel electrophoresis) and were then transferred to PVDF membranes (Bio-Rad, Hercules, CA, USA). The electroblotted membranes were blocked by TBS containing 5% non-fat milk (Santa Cruz, Dallas, TX, USA) and were probed with primary antibodies overnight at 4°C and immunoblotted with specific antibodies. Primary antibodies against the following proteins were used: UCP1, PGC1α, PRDM16, Cidea (Abcam, Cambridge, UK), PPARγ (CST, Danvers, MA, USA), 422/Ap2 (Santa Cruz Biotechnology), RPS3A (ProteinTech Group, Rosemont, IL, USA), Cyto c (CST), Actin (Sigma, St. Louis, USA).
Rps3a
Manufactured by Proteintech
Sourced in United States
RPS3A is a laboratory equipment product offered by Proteintech. It serves as a ribosomal protein, functioning as a component of the small ribosomal subunit.
Automatically generated - may contain errors
Lab products found in correlation
Cocktail of proteinase inhibitors, by Roche (1 mentions)
Cidea, by Abcam (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Tsg101 sc 7964, by Santa Cruz Biotechnology (1 mentions)
Flotillin 1, by BD (1 mentions)
Rpl24, by Proteintech (1 mentions)
Ecl kit, by Merck Group (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
Polyvinylidene fluoride membrane, by Bio-Rad (1 mentions)
β actin, by Abcam (1 mentions)
2 protocols using rps3a
1
Protein Expression Analysis in Cultured Cells
2
Exosomal Protein Profiling by Western Blot
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Samples containing 15 μg of exosomal proteins derived from the serum or cell lysates of a study subject were denatured with SDS (Sigma) loading buffer, boiled for up to 5 min, separated by SDS-PAGE, and transferred onto a polyvinylidene fluoride membrane (Bio-Rad) at 30 V for 60 min. Next, the membranes were blocked with 5% nonfat dry milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) at room temperature for 1 h and subsequently incubated overnight at 4 °C with primary antibodies against TSG101 (sc-7964; Santa Cruz Biotechnology), HSP70 (4873S; CST), Alix (2171S; CST), CD63 (ab59479; Abcam), Calreticulin (12238T; CST), CD151 (96282S; CST), ITGB1 (4706S; CST), Flotillin-1 (610820; BD), β-actin (ab3280; Abcam), RPL6 (15387; Proteintech), RPL13 (11271; Proteintech), RPL24 (17082; Proteintech), RPS3A (14123; Proteintech), RPS8 (18228; Proteintech), RPS10 (14894; Proteintech), C3 (21337; Proteintech), C5 (66634; Proteintech), and C7 (17642; Proteintech). The membranes were washed four times with TBST and incubated with horse radish peroxide–conjugated secondary antibodies at 37 °C for 1 h. Finally, the membranes were washed with TBST four additional times, after which immunoreactive bands were detected with an ECL Kit (Millipore).
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