Total protein of quadriceps muscle was harvested and lysed in buffer containing complete protease inhibitor cocktail (Sigma) as described previously.3 (link) Muscle tissue cytosolic and nuclear proteins were separated by using a NE-PER kit (Pierce Biotechnology, Rockford, IL, USA) according to the manufacturer’s instructions. The protein concentration was determined by BCA Protein Assay (Pierce Biotechnology) and equal amount of total protein were loaded to sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel for Western blotting analysis described previously.21 (link) The following primary antibodies were used in the analysis: anti-STARS (1:1,000; Abcam), anti-RhoA (1:2,000; Abcam), anti-SRF (1:1,000; Abcam), anti-histone H3 (1:1,000; Abcam), anti-glyceraldehyde 3-phosphate dehydrogenase (1:30,000; CST), and anti-β-tubulin (1:30,000; Sigma).
Anti srf
Manufactured by Abcam
Anti-SRF is a laboratory reagent that can be used to detect and quantify the presence of the Serum Response Factor (SRF) protein in biological samples. SRF is a transcription factor that plays a critical role in regulating gene expression, particularly in processes related to cell growth, differentiation, and development. Anti-SRF can be utilized in various research applications, such as Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA), to investigate the expression and distribution of SRF in different cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Ne per kit, by Thermo Fisher Scientific (1 mentions)
Anti rhoa, by Abcam (1 mentions)
Anti histone h3, by Abcam (1 mentions)
Anti β tubulin, by Merck Group (1 mentions)
Complete protease inhibitor cocktail, by Merck Group (1 mentions)
Bca protein assay, by Thermo Fisher Scientific (1 mentions)
Anti sirt1, by Cell Signaling Technology (1 mentions)
Anti pnf bp65, by Cell Signaling Technology (1 mentions)
Anti nrf2, by Abcam (1 mentions)
Anti trx2, by Proteintech (1 mentions)
Anti srf, by Proteintech (1 mentions)
Anti sod2, by Proteintech (1 mentions)
Anti e2f2, by Abcam (1 mentions)
Anti β mhc, by Abcam (1 mentions)
Anti anp, by Abcam (1 mentions)
Anti nf bp65, by Cell Signaling Technology (1 mentions)
Anti α sma, by Abcam (1 mentions)
Anti col1a1, by Proteintech (1 mentions)
Anti ho 1, by Proteintech (1 mentions)
Anti pgc 1α, by Abcam (1 mentions)
3 protocols using anti srf
1
Quadriceps Muscle Protein Analysis
2
Comprehensive Western Blot Analysis
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Western blot assays were performed as described previously,39 (link) using anti-ANP, anti-β-MHC, anti-α-SMA, anti-p-Smad1, anti-Smad1, anti-SRF, anti-E2F2, anti-PGC-1α, anti-NRF2 (Abcam), anti-ACTA1, anti-COL1A1, anti-COL3A1, anti-TRX2, anti-SOD2, anti-HO-1, anti-SRF (Proteintech), anti-SIRT1, anti-p-Smad3, anti-Smad3, anti-p-NF-кB p65, anti-NF-кB p65 (Cell Signaling Technology), anti-RB1or anti-GAPDH (Santa Cruz Biotechnology).
3
Evaluating Biomarkers in Glioblastoma Tissues
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Archival FFPE GBM tumor tissues from a consecutive cohort of 286 patients treated in the UMCU between 2009 and 2013 were included on tissue microarrays (TMA’s) as described previously [7 (link)]. Immunohistochemistry was performed, as described previously [7 (link)], with antibodies against STAT5b (phospho S731, Rabbit polyclonal, Abcam, Cambridge, UK), VEGF (Rabbit polyclonal, ThermoScientific, Waltham, MA USA), anti-NF-κB p65 (phospho S276, Rabbit polyclonal, Abcam); anti-STAT3 (phospho Y705) (Rabbit monoclonal, Cell Signaling, Leiden, The Netherlands); anti-CEBP-β (Mouse monoclonal, Abcam); anti-SRF (Rabbit polyclonal, Abcam, and rabbit polyclonal, Sigma-Aldrich, St. Louis, MO, USA).
Protein expression evaluation was performed with blinding for the clinical data, and was supervised by a senior neuropathologist. The percentage of nuclear and/or cytoplasmatic staining was scored as: 0, negative; 1, 1–25% positive cells; 2, 26–50% positive cells; 3, 51–75% positive cells and 4, 76–100% positive cells. A mean expression score was computed per patient. Due to insufficient tissue quality on TMA, a variable number of tissues per staining could not be evaluated (n = 8–21).
Protein expression evaluation was performed with blinding for the clinical data, and was supervised by a senior neuropathologist. The percentage of nuclear and/or cytoplasmatic staining was scored as: 0, negative; 1, 1–25% positive cells; 2, 26–50% positive cells; 3, 51–75% positive cells and 4, 76–100% positive cells. A mean expression score was computed per patient. Due to insufficient tissue quality on TMA, a variable number of tissues per staining could not be evaluated (n = 8–21).
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