Cells were lysed with RIPA lysis buffer. After centrifugation at 10,000× g for 15 min, supernatants were harvested for SDS-PAGE and Western blotting. Protein quantification was performed by the BCA assay (Sigma). Proteins (40 μg) were separated on SDS-PAGE and transferred onto PVDF membranes. Membranes were then blocked and probed using the following primary antibodies: rabbit anti-MCU (1:1000; Cell Signaling, Danvers, MA, USA) and mouse anti-GAPDH at 1:5000 (Santa Cruz, Santa Cruz, CA, USA). After washing, membranes were incubated with appropriate secondary HRP-conjugated antibodies (Jackson ImmunoResearch, Bar Harbor, ME, USA) at 1:10,000. Bands were detected by chemiluminescence using the LiteAblot Turbo (Euroclone, Milan, Italy). Images were acquired with Image Quant Las 4000 (GE Healthcare, Chicago, IL, USA). Densitometric analysis was performed with Image J 1.47v (NIH, USA).
Rabbit anti mcu
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit anti-MCU is a primary antibody designed to detect the Mitochondrial Calcium Uniporter (MCU) protein. MCU is a key component of the mitochondrial calcium uptake machinery. This antibody can be used in various applications such as Western blotting, immunoprecipitation, and immunohistochemistry to study the expression and localization of MCU in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Imagequant las 4000, by GE Healthcare (1 mentions)
Mouse anti gapdh, by Santa Cruz Biotechnology (1 mentions)
Bca assay, by Merck Group (1 mentions)
Liteablot turbo, by Euroclone (1 mentions)
Hrp conjugated antibodies, by Jackson ImmunoResearch (1 mentions)
Las 4000, by Cytiva (1 mentions)
Rabbit anti akt, by Cell Signaling Technology (1 mentions)
Mouse anti flag, by Abmart (1 mentions)
Rabbit anti ha, by Cell Signaling Technology (1 mentions)
Rabbit anti micu1, by Merck Group (1 mentions)
Ab10983, by Abcam (1 mentions)
Mouse anti hsp90α β, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti akt ps473, by Cell Signaling Technology (1 mentions)
2 protocols using rabbit anti mcu
1
Protein Expression Analysis by Western Blot
2
Western Blot Analysis of Mitochondrial Proteins
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Tissues were lysed in modified RIPA buffer (450 mM NaCl, 1% NP-40, 0.1% SDS, 0.5% Deoxycholic acid (sodium salt), 50 mM Tris pH 7.5, 1x complete protease inhibitor). Total cellular protein (30 μg) was separated on SDS-PAGE gels and transferred to nitrocellulose membrane. After blocking in 5% defatted milk, membranes were incubated in primary antibody at 4°C overnight. The primary antibodies used in this study were: rabbit anti-UCP1 (Abcam, ab10983, 1:40,000 for BAT, 1:2000 for co-IP experiments), rabbit anti-EMRE (Santa Cruz Biotech, sc-86337, 1:300), rabbit anti-MCU (Cell Signaling Technologies, 14997, 1:2000) , rabbit anti-MICU1 (Sigma, HPA037479, 1:1000), rabbit anti-HA (Cell Signaling Technologies, 3724, 1:2000) , mouse anti-SDHB (Santa Cruz Biotech, sc-271548, 1:2000) , mouse anti-FLAG (Abmart, 314375, 1:5000), mouse anti-PDHE1α (Santa Cruz Biotech, sc-377092, 1:5000), rabbit anti-phospho-PDHE1α S293 (Millipore, ABS204, 1:2000), rabbit anti-AKT(pS473) (Cell Signaling Technologies, 9271, 1:2000) , rabbit anti-AKT (Cell Signaling Technologies, 9272, 1:2000) , mouse anti-HSP90α/β (Santa Cruz Biotech, sc13119, 1:10000), mouse anti-NDUFS1 (Abcam, ab22094, 1:10000). The membranes were then incubated in HRP-conjugated secondary antibodies at 37°C for 1 h and visualized using Tanon 5200 Chemiluminescent Imaging System.
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