Trilobatin, palmitate, DAPI and (2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino)-2-deoxyglucose (2NBDG) uptake measurement kits were purchased from Sigma (St. Louis, MO, USA). IRS1, p-IRS1 (Ser 612), p-IRS1 (Ser 307), Akt, p-Akt (Ser 473), p-Akt (thr308), Na, K-ATPase, MYH1, MYOD1, and β-actin primary antibodies were bought from Cell Signaling Technology (Danvers, MA, USA). GLUT4 antibody was obtained from Abcam (Cambridge, MA, USA). HRP-conjugated GAPDH primary antibody was purchased from Aksmics (shanghai, China), Specific anti-mouse and anti-rabbit HRP-conjugated second antibodies were obtained from Santa Cruz Biotechnology (Texas, CA, USA). Rat/mouse insulin ELISA kits (EZRMI-13K) and ECL chemiluminescence detection reagent were obtained from Millipore (Billerica, MA, USA). Plasma membrane protein extraction kit, nuclear/cytosolic fractionation kit, RIPA buffer and BCA protein assay kit and other chemicals were purchased from Beyotime (Shanghai, China).
Nuclear cytosolic fractionation kit
Manufactured by Beyotime
Sourced in China
The Nuclear/Cytosolic Fractionation Kit is a laboratory tool designed to separate the nuclear and cytosolic fractions of cells. It provides a straightforward method to isolate these cellular components for further analysis and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Irs 1, by Cell Signaling Technology (3 mentions)
Trilobatin, by Merck Group (3 mentions)
Myod1, by Cell Signaling Technology (3 mentions)
Rat mouse insulin elisa kits ezrmi 13k, by Merck Group (3 mentions)
Ecl chemiluminescence detection reagent, by Merck Group (3 mentions)
Plasma membrane protein extraction kit, by Beyotime (3 mentions)
Glut4 antibody, by Abcam (3 mentions)
β actin, by Cell Signaling Technology (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions)
Palmitate, by Merck Group (3 mentions)
Ripa buffer, by Beyotime (3 mentions)
Bca protein assay kit, by Beyotime (3 mentions)
Speci c anti mouse and anti rabbit hrp conjugated second antibodies, by Santa Cruz Biotechnology (2 mentions)
Specific anti mouse and anti rabbit hrp conjugated second antibodies, by Santa Cruz Biotechnology (1 mentions)
5 protocols using nuclear cytosolic fractionation kit
1
Glucose Uptake and Insulin Signaling Pathway
2
Subcellular Fractionation of IEC-6 Cells
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Subfractionation was performed using a Nuclear/Cytosolic Fractionation Kit (Beyotime, Wuhan, China). IEC-6 cultures were washed with ice-cold PBS, scraped from the plates, and collected. After centrifugation, the supernatant was discarded, and the cells were suspended with Cytosol Extraction Buffer containing DTT/protease inhibitors, incubated on ice for 10 min, and Cell Lysis Reagent was added. The nuclei fraction was fractioned at 800 × g for 10 min. The supernatant was further centrifuged at 12 000 × g for 10 min, and the final supernatant was collected for cytoplasmic fraction. The nuclei pellet was washed and resuspended with Nuclear Extraction Buffer containing DTT/protease inhibitors.
3
Insulin Signaling and Glucose Uptake Assay
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Trilobatin, palmitate, DAPI and (2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino)-2-deoxyglucose (2NBDG) uptake measurement kits were purchased from Sigma (St Louis, MO, USA). IRS1, p-IRS1 (Ser 612), p-IRS1 (Ser 307), Akt, p-Akt (Ser 473), p-Akt (thr308), Na,K-ATPase, MYH1, MYOD1, and β-actin primary antibodies were bought from Cell Signaling Technology (Danvers, MA, USA). GLUT4 antibody was obtained from Abcam (Cambridge, MA, USA). HRP-conjugated GAPDH primary antibody was purchased from Aksmics (shanghai, China), Speci c anti-mouse and anti-rabbit HRP-conjugated second antibodies were obtained from Santa Cruz Biotechnology (Texas, CA, USA). Rat/mouse insulin ELISA kits (EZRMI-13K) and ECL chemiluminescence detection reagent were obtained from Millipore (Billerica, MA, USA). Plasma membrane protein extraction kit, nuclear/cytosolic fractionation kit, RIPA buffer and BCA protein assay kit and other chemicals were purchased from Beyotime (Shanghai, China).
4
Glucose Uptake and Insulin Signaling
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Trilobatin, palmitate, DAPI and (2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino)-2-deoxyglucose (2NBDG) uptake measurement kits were purchased from Sigma (St Louis, MO, USA). IRS1, p-IRS1 (Ser 612), p-IRS1 (Ser 307), Akt, p-Akt (Ser 473), p-Akt (thr308), Na,K-ATPase, MYH1, MYOD1, and β-actin primary antibodies were bought from Cell Signaling Technology (Danvers, MA, USA). GLUT4 antibody was obtained from Abcam (Cambridge, MA, USA). HRP-conjugated GAPDH primary antibody was purchased from Aksmics (shanghai, China), Speci c anti-mouse and anti-rabbit HRP-conjugated second antibodies were obtained from Santa Cruz Biotechnology (Texas, CA, USA). Rat/mouse insulin ELISA kits (EZRMI-13K) and ECL chemiluminescence detection reagent were obtained from Millipore (Billerica, MA, USA).
Plasma membrane protein extraction kit, nuclear/cytosolic fractionation kit, RIPA buffer and BCA protein assay kit and other chemicals were purchased from Beyotime (Shanghai, China).
Plasma membrane protein extraction kit, nuclear/cytosolic fractionation kit, RIPA buffer and BCA protein assay kit and other chemicals were purchased from Beyotime (Shanghai, China).
5
Subcellular Protein Fractionation Protocol
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To measure the expression of BAX, cytochrome C, and cleaved PARP in the mitochondria and the cytosol, mitochondrial and cytosolic proteins were isolated using a mitochondrial/cytosol fractionation kit (Beyotime Biotechnology, Inc., Beijing, China). The cytosolic and nuclear proteins were obtained using a nuclear/cytosolic fractionation kit (Beyotime Biotechnology, Inc., Beijing, China). In brief, cells were collected by trypsinization at the indicated times, washed with PBS, incubated with ice-cold cytosolic separation buffer, agitated and lysed on ice for 15 min. The cytosolic, nuclear and mitochondrial fractions were then harvested by a series of centrifugation steps in accordance with the manufacturer's instructions.
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