Cells were lysed in ice-cold RIPA cell buffer (Sigma) supplemented with protease inhibitors (Sigma). The proteins were separated with a 4–12% PAGE gel and electrotransferred onto a PVDF membrane. The membrane was probed with primary antibodies and subsequently detected by horseradish-peroxidase (HRP) conjugated antibodies (Santa Cruz). The primary antibodies were β-catenin (Santa Cruz), Flag antibody (Sigma), HIF1α (Santa Cruz) and HIF2α (Santa Cruz).
Ripa cell buffer
Manufactured by Merck Group
Sourced in United States
RIPA cell buffer is a lysis buffer used for the extraction of proteins from cells. It is a detergent-based buffer that aids in the disruption of cell membranes and the solubilization of proteins. The buffer contains a combination of ionic and non-ionic detergents, as well as other components such as salts and buffers, to ensure efficient protein extraction and preservation.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase hrp conjugated antibody, by Santa Cruz Biotechnology (1 mentions)
Hif 2α, by Santa Cruz Biotechnology (1 mentions)
Flag antibody, by Merck Group (1 mentions)
β catenin, by Santa Cruz Biotechnology (1 mentions)
Hif 1α, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor, by Merck Group (1 mentions)
α tubulin, by Santa Cruz Biotechnology (1 mentions)
2 protocols using ripa cell buffer
1
Protein Extraction and Western Blot Analysis
2
Protein Expression and Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were lysed in ice-cold RIPA cell buffer (Sigma, USA) supplemented with protease inhibitors (TransGen Biotech, China). Proteins were separated on a 10% PAGE gel (made in-house) and electrotransferred onto a PVDF membrane. Probing was performed with specific primary antibodies and HRP-conjugated secondary antibodies. The primary antibodies used were HA (3724S, Cell Signaling Technology, 1:1000), Flag (M2, Sigma, 1:1000) and α-Tubulin (SC-8035, Santa Cruz Biotechnology, 1:1000).
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