H9c2 cells were lysed in RIPA lysis buffer (Invitrogen, Carlsbad, CA, United States), supplemented with 1 mM PMSF and 1% protease inhibitor cocktail (Sigma-Aldrich). Lysates were incubated on ice for 30 min followed by centrifugation at 17,000 g for 15 min at 4°C. The supernatant was collected and quantified using the bicinchoninic acid assay (Thermo Fisher Scientific). Proteins were resolved on 4–20% mini-PROTEAN TGX gels (BioRad) and transferred to nitrocellulose membranes (0.45 μm, BioRad). The membranes were blotted with the following antibodies: Letm1 (16024-1-AP; Proteintech), NCLX (ab83551; Abcam), MCU (14997; Cell Signaling Technology) and GAPDH (CB1001; Millipore). GAPDH was used as the loading control. The LI-COR infrared fluorescent mouse (925–68020) and rabbit (925–32211) secondary antibodies were used for visualization by a LI-COR Odyssey scanner. Band quantification (densitometry) was performed using ImageJ and Origin 8 software (OriginLab Corporation, Northampton, MA, United States).
Letm1 16024 1 ap
Manufactured by Proteintech
Letm1 (16024-1-AP) is a primary antibody product offered by Proteintech. It is used for detecting the Letm1 (leucine zipper-EF-hand containing transmembrane protein 1) protein in various applications. The product is intended for research use only.
Automatically generated - may contain errors
Lab products found in correlation
Ripa lysis buffer, by Thermo Fisher Scientific (1 mentions)
Bicinchoninic acid assay, by Thermo Fisher Scientific (1 mentions)
Origin 8, by OriginLab (1 mentions)
Odyssey scanner, by LI COR (1 mentions)
Imagej, by OriginLab (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Mini protean tgx gel, by Bio-Rad (1 mentions)
Nitrocellulose membranes 0.45 μm, by Bio-Rad (1 mentions)
Mcu 14997, by Cell Signaling Technology (1 mentions)
Irdye 680lt goat anti mouse igg, by LI COR (1 mentions)
Gapdh 2118, by Cell Signaling Technology (1 mentions)
Vdac ab14734, by Abcam (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Irdye 800cw goat anti rabbit immunoglobulin g, by LI COR (1 mentions)
Sdha ab14715, by Abcam (1 mentions)
2 protocols using letm1 16024 1 ap
1
Protein Analysis of H9c2 Cell Lysates
2
Immunoblotting Analysis of Metabolic Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Specific proteins were detected using the following rabbit monoclonal antibodies: phospho-SAPK/JNK (Thr183/Tyr185, 4668), SAPK/JNK (9252), phospho-Akt (Ser473, 4060), Akt (9272), S6 ribosomal protein (2217S), phospho–S6 ribosomal protein (4856S), glyceraldehyde phosphate dehydrogenase (GAPDH) (2118), LC3A/B (12741) MCU (14997) (Cell Signaling Technologies), and MRPP3 (HPA020459) (Sigma-Prestige) diluted 1:1000; rabbit polyclonal antibodies: GLUT2 (54460) (Abcam) diluted 1:1000, GLUT4 (PA-1-1065) (Thermo Fisher Scientific), CPTII (OAAN00972) (Aviva System Biology) diluted 1:500, Letm1 (16024-1-AP) (Proteintech) diluted 1:1000, MRPP2 (HPA001432) (Sigma-Prestige) diluted 1:1000, LRPPRC (sc66844) (Santa Cruz Biotechnology) diluted 1:500, TFAM (ab131607) (Abcam) diluted 1:1000; mouse monoclonal antibodies: total OXPHOS Cocktail Antibody (ab110412), VDAC (ab14734), and SDHA (ab14715) (Abcam), diluted 1:1000. IRDye 800CW goat anti-rabbit immunoglobulin G (IgG) or IRDye 680LT goat anti-mouse IgG (LI-COR Biosciences) secondary antibodies were used, and the immunoblots were visualized using an Odyssey infrared imaging system (LI-COR Biosciences). Relative amount of protein was determined using the Image Studio Lite Software.
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!