Rapid-immunoprecipitation assay (RIPA) buffer, blocking solution, and protease inhibitor were purchased from GenDepot. Anti-RUNX2 and -GAPDH antibodies were purchased from Detroit R&D (Detroit, MI). Anti-DLX5 and -PHEX were purchased from Proteintech (Rosemont, IL, USA). Anti-OCN and -OSX were purchased from Abcam (Cambridge, UK). Cell lysates were prepared from fusion bed tissue using RIPA buffer. Proteins were resolved on SDS-PAGE and transferred to 0.45 μm nitrocellulose blotting membranes (Amersham Protran, Germany). The blots were blocked using superblocking solution (Gengepot) for 1 hour at room temperature and probed with primary antibody overnight at 4°C. Blots were washed with PBST and incubated with an anti-rabbit or –mouse peroxidase-conjugated secondary antibody for 1 hour at room temperature. Signal was visualized by enhanced chemiluminescence (ECL) using Azure300 (Azure Biosystems, Dublin, CA, USA), and intensities were quantified using a computing densitometry program from Image Studio Lite (LI-COR, Lincoln, NE, USA).
Rapid immunoprecipitation assay ripa buffer
Manufactured by GenDEPOT
Sourced in United States
RIPA buffer is a solution used in molecular biology and biochemistry to extract and solubilize proteins from cells and tissues. It is a detergent-based buffer that disrupts cell membranes and denatures proteins, allowing for the extraction and purification of proteins from samples.
Automatically generated - may contain errors
Lab products found in correlation
Image studio lite, by LI COR (2 mentions)
Azure 300, by Azure Biosystems (2 mentions)
Blocking solution, by GenDEPOT (2 mentions)
Protease inhibitor, by GenDEPOT (2 mentions)
Protran, by Cytiva (1 mentions)
Total erk1 2, by Cell Signaling Technology (1 mentions)
Total p38, by Cell Signaling Technology (1 mentions)
Integrin α5, by Santa Cruz Biotechnology (1 mentions)
Integrin β1, by Santa Cruz Biotechnology (1 mentions)
P erk1 2, by Cell Signaling Technology (1 mentions)
Integrin αv, by Santa Cruz Biotechnology (1 mentions)
N cadherin, by Santa Cruz Biotechnology (1 mentions)
E cadherin, by Santa Cruz Biotechnology (1 mentions)
2 protocols using rapid immunoprecipitation assay ripa buffer
1
Western Blot Analysis of Osteogenic Markers
2
Western Blot Analysis of Integrin and Cadherin Signaling
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Rapid-immunoprecipitation assay (RIPA) buffer, blocking solution, and protease inhibitor were purchased from GenDepot (Barker, TX, USA). β-Tubulin and RUNX2 antibodies were purchased from Proteintech (Rosemont, IL, USA), and integrin α5, integrin αV, N-cadherin, E-cadherin, and integrin β1 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Total p38, p–p38, total ERK1/2, and p–ERK1/2 antibodies were purchased from Cell Signaling Technology (Billerica, MA, USA). After primary antibody incubation, membranes were washed with PBST and incubated with horseradish conjugated secondary antibodies for 1 h at room temperature. Signal was visualized by enhanced chemiluminescence (ECL) using Azure300 (Azure Biosystems, Dublin, CA, USA), and intensities were quantified using a computing densitometry program from Image Studio Lite (LI-COR, Lincoln, NE, USA).
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