Cells were seeded overnight in 6 well plates and then treated for the designated period. Total cellular protein was extracted using RIPA buffer and the protein concentration was determined by using Pierce BCA protein assay kit (ThermoFisher, USA). Equal amounts of proteins were loaded onto a polyacrylamide gel and separated by SDS-PAGE. The proteins were transferred overnight to a nitrocellulose membrane (Biorad, USA). The membranes were blocked for 1h using membrane blocking solution (Life Technologies, USA) and then incubated overnight with primary antibody at 4°C. The primary antibodies used were rabbit monoclonal anti-Id1 (Calbioreagents, San Mateo, CA), rabbit monoclonal XIAP, rabbit monoclonal beta III tubulin (Tuj1), rabbit monoclonal anti-pSmad 1/5 (Cell Signaling Technology, MA, USA), rabbit polyclonal anti-Smad 1/5 (Upstate Biotechnology, NY, USA), rabbit anti-actin, rabbit polyclonal anti-GAPDH (Sigma, St. Louis, MO), and mouse monoclonal anti-spectrin (EMD Millipore, CA, USA). Rabbit polyclonal SOX2 (Abcam, USA), mouse monoclonal anti-NeuN (Chemicon, USA), and mouse monoclonal anti-Nestin (Chemicon, USA).
Rabbit monoclonal xiap
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit monoclonal XIAP is a lab equipment product manufactured by Cell Signaling Technology. It is an antibody that recognizes the X-linked inhibitor of apoptosis protein (XIAP).
Automatically generated - may contain errors
Lab products found in correlation
Rabbit monoclonal anti psmad1 5, by Cell Signaling Technology (2 mentions)
Rabbit anti actin, by Merck Group (2 mentions)
Membrane blocking solution, by Thermo Fisher Scientific (1 mentions)
Mouse monoclonal anti spectrin, by Merck Group (1 mentions)
Mouse monoclonal anti neun, by Merck Group (1 mentions)
Mouse monoclonal anti nestin, by Merck Group (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Enhanced chemiluminescence system, by Cytiva (1 mentions)
Rabbit monoclonal anti ptak1, by Cell Signaling Technology (1 mentions)
Rabbit monoclonal anti activated caspase 3, by Cell Signaling Technology (1 mentions)
Polyclonal rabbit anti gapdh, by Merck Group (1 mentions)
2 protocols using rabbit monoclonal xiap
1
Western Blot Protein Analysis Protocol
2
Protein Extraction and Western Blot Analysis
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Total cellular protein was prepared using RIPA buffer containing a protease inhibitor cocktail and protein concentration was measured using the BCA assay as described [4 (link)]. In brief, protein was analyzed by SDS-PAGE, transferred to nitrocellulose (Schleicher and Schuell, Keene, NH). After blocking, the blots were incubated overnight at 4 °C with the appropriate primary antibody in Tris-buffered saline with 1 % Tween (TBST) and 5 % BSA. Secondary antibodies were applied for 1 h at room temperature. Specific proteins were detected using the enhanced chemiluminescence system (Amersham, Arlington Heights, IL). The primary antibodies that were used were rabbit monoclonal anti-pSmad 1/5, rabbit monoclonal anti-pSmad2, rabbit monoclonal anti-Smad2, rabbit monoclonal anti-pTAK1, rabbit monoclonal XIAP, rabbit anti-monoclonal p-p65, rabbit monoclonal anti-activated caspase-3, rabbit polyclonal anti-BMPRII, and rabbit monoclonal anti-EGR-1 (Cell signaling Technology, Danvers MA), Rabbit monoclonal anti-TAK1 (Invitrogen, Grand Island NY), rabbit anti-actin, an affinity isolated antigen specific antibody (Sigma, Saint Louis, MO), rabbit monoclonal anti-Id1 and rabbit monoclonal anti-Id3 (Calbioreagents, San Mateo, CA), rabbit polyclonal anti-GAPDH (Sigma, St. Louis, MO) and rabbit polyclonal pMEK-1/2 (Cell Signaling, Danvers MA).
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