Cells were lysed in Cell Lysis Buffer (Cell Signaling), supplemented with protease (#11697498001; Roche) and phosphatase inhibitors (#04906845001; Roche), in addition to 0.2 M glycerol-2-phosphate (Sigma-Aldrich) and resolved on NuPAGE gels (Novex; Life Technologies). Primary antibodies used in this study are β-actin (1:10,000, Cat. #A5441; Sigma-Aldrich), CAP350 (1:1000, Cat. #20022-1-AP; Acris Antibodies, San Diego, CA, USA), FLAG-TRα (1:1000, anti-FLAG #F7425; Sigma-Aldrich), GAPDH (1:10,000, Cat. #G8795; Sigma-Aldrich), MuRF1 (1:250–1:1000, #AF5366; R&D Systems), GAPDH (1:4000, G8795; Sigma-Aldrich), p-Rb (#sc-50; Santa Cruz Biotechnology), myc-tagged MuRF1 (1:4000, anti-myc, #C4439; or anti-myc-HRP-linked primary, 1:10,000, #A5598; Sigma-Aldrich), glutathione-S-transferase (GST)-tagged TRα (anti-GST; #G1160; Sigma-Aldrich, 1:10,000), TRα (PA1-211A; Thermo Scientific, 1:500), and HA-ubiquitin (anti-HA; #sc-57592, 1:1000). HRP-linked secondary antibodies used in this study are anti-mouse (#NA931V; GE Healthcare Life Sciences, 1:10,000), anti-rabbit (#A9169, Sigma-Aldrich, 1:20,000), and anti-goat (#A5420; Sigma-Aldrich, 1:10,000).
Pa1 211a
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PA1-211A is a laboratory pipette designed for precise fluid handling. It features adjustable volume settings and is suitable for a variety of laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Nupage gel, by Thermo Fisher Scientific (1 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
Glycerol 2 phosphate, by Merck Group (1 mentions)
Na931v, by GE Healthcare (1 mentions)
Murf1, by R&D Systems (1 mentions)
Anti gst, by Merck Group (1 mentions)
Gapdh, by Merck Group (1 mentions)
Anti flag, by Merck Group (1 mentions)
Phosphatase inhibitor, by Roche (1 mentions)
Ab3625, by Abcam (1 mentions)
Ab180612, by Abcam (1 mentions)
Criterion tgx precast gel, by Bio-Rad (1 mentions)
Ma3 922, by Thermo Fisher Scientific (1 mentions)
Trans blot turbo, by Bio-Rad (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Ab8245, by Abcam (1 mentions)
2 protocols using pa1 211a
1
Assaying Protein Interactions and Modifications
2
Immunoblotting Analysis of Cardiac Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunoblotting was performed as previously described.38 Briefly, protein extracts from the heart were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using 4-20% gradient polyacrylamide gels (Criterion™ TGX™ Precast Gels, No. 5671095; Bio-Rad) and then electroblotted into nitrocellulose membranes (Trans-Blot® Turbo; Bio-Rad). Blots were blocked and incubated with primary antibodies to phospholambam (MA3-922; Thermo Fisher Scientific), phospho-phospholambam (8496; Cell Signaling) ATPase sarcoplasmic/endoplasmic reticulum Ca2+ transporting 2 (SERCA2a; PA1-211A/IRDye 800; ab3625; Abcam), and thyroid hormone receptors a (PA1-211A; Thermo Fisher Scientific) and b (ab180612; Abcam) overnight at 4ºC. The immunoblots were subsequently washed and incubated with 700 or 800 nm infrared dye-conjugated antibodies (Nos. 926-68020 and 926-32211; LI-COR Biosciences). The membrane was imaged by scanning at 800 and 700 nm with an Odyssey Infrared Imaging System (LI-COR Biosciences). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control (ab8245; Abcam) and the control group was set as reference.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!