Cells were lysed with RIPA buffer (Thermo Fisher Scientific) in the presence of a protease inhibitor and phosphatase inhibitor cocktail (Thermo Fisher Scientific). Lysates were separated by SDS–PAGE under reducing conditions, transferred to a nitrocellulose membrane, and analyzed by immunoblotting. MDHC (H-6) (Cat# sc-166879, RRID:AB_10609257) was purchased from Santa Cruz Biotechnology. GPD1 polyclonal antibody (Cat# 27943-1-AP, RRID:AB_2881016) and GPD1L polyclonal antibody (Cat# 17263-1-AP, RRID:AB_2112359) were purchased from Proteintech. Rabbit anti-Ras (G12V) monoclonal antibody (Cat.#: 14412, RRID:AB_2714031) was purchased from Cell Signaling. Goat anti-rabbit secondary antibodies (HRP conjugated) and anti-mouse secondary antibodies were purchased from LI-COR and Santa Cruz Biotechnology, respectively. Immunoblotting for β-tubulin by rabbit anti-β-tubulin antibody (HRP conjugated) (Cat# 5346, RRID:AB_1950376) (Cell Signaling) was used as a loading control. Signal was detected by using WesternSure premium chemiluminescent substrate and the C-Digit Blot Scanner (LI-COR) according to the manufacturer’s instructions.
Anti mouse secondary antibodies
Manufactured by LI COR
Anti-mouse secondary antibodies are laboratory reagents used to detect and visualize primary antibodies that have been raised against mouse antigens. These secondary antibodies are designed to specifically bind to the Fc (constant) region of mouse primary antibodies, allowing for the indirect detection and analysis of target proteins or molecules in various immunoassays and immunohistochemical applications.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Protease inhibitor and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
C digit blot scanner, by LI COR (1 mentions)
Westernsure premium chemiluminescent substrate, by LI COR (1 mentions)
Odyssey infrared imaging system, by Gene Tech (1 mentions)
2 protocols using anti mouse secondary antibodies
1
Immunoblotting of Protein Lysates
2
Investigating RXRα and ERα Interaction
Check if the same lab product or an alternative is used in the 5 most similar protocols
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HUVEC cells were co-transfected with RXRα and ERα. Protein extracts were isolated from cells using RIPA buffer. Tagged proteins were immunoprecipitated overnight at 4 °C using protein A/G agarose (CW0349) and anti-RXRα antibody, and then all complexes were pelleted at 3000 rpm for 3 min. The beads were washed and fractionated by 12% SDS-PAGE, followed by transfer to a nitrocellulose membrane. The membrane was immunoblotted with mouse anti-ERα (1:5000) and β-actin (1:250) overnight at 4 °C and then incubated with anti-mouse secondary antibodies (Li-COR Biosciences) for 1 h at RT. The specific proteins were visualized by Odyssey Infrared Imaging System (Gene Company Limited).
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