Protein contents were extracted with RIPA lysis buffer (P0013B; Beyotime) supplemented with 1% proteinase inhibitor phenyl‐methylsulfonyl fluoride (PMSF; ST506‐2; Beyotime). Isolated target protein on SDS‐PAGE gels were transferred onto PVDF membranes (IPVH00010; Millipore). The blotting membranes were treated with 5% skim milk for 1 h and then with primary antibody overnight (12–16 h). Thereafter, blotting bands were incubated with secondary antibodies at room temperature for 2 h before visualization with Immobilon® Western (P90719, Millipore). In this study, primary antibodies (1:1000) included: anti‐β‐actin (200068‐8F10; ZEN‐BIO), anti‐AFF4 (14662‐1‐AP; Proteintech), anti‐LC3B (18725‐1‐AP; Proteintech), anti‐Collagen I (R26615; ZEN‐BIO), anti‐Runx2 (860139; ZEN‐BIO), anti‐ALP (381009; ZEN‐BIO), anti‐ULK1(381887; ZEN‐BIO), anti‐Phospho‐ULK1 (Ser758) (AF4387; Affinity Biosciences), anti‐mTOR (380411; ZEN‐BIO), anti‐Phospho‐mTOR (Ser2448) (381557; ZEN‐BIO). Secondary antibodies included: Goat anti‐mouse IgG (H&L) (HRP conjugated) (511103; ZEN‐BIO) and Goat anti‐rabbit IgG (H&L) (HRP conjugated) (511203, ZEN‐BIO).
Anti mtor
Manufactured by ZenBio
Sourced in Germany, United Kingdom
Anti-mTOR is a laboratory product that functions as an inhibitor of the mammalian target of rapamycin (mTOR) protein. mTOR is a key regulator of cell growth, proliferation, and metabolism. Anti-mTOR is used in research settings to study the role of mTOR signaling in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti p mtor, by ZenBio (2 mentions)
St506 2, by Beyotime (1 mentions)
Immobilon western, by Merck Group (1 mentions)
Anti β actin 200068 8f10, by ZenBio (1 mentions)
Anti aff4 14662 1 ap, by Proteintech (1 mentions)
Anti collagen 1, by ZenBio (1 mentions)
Anti runx2, by ZenBio (1 mentions)
Anti alp, by ZenBio (1 mentions)
Ipvh00010, by Merck Group (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti caspase 9, by Proteintech (1 mentions)
Anti bcl 2, by Proteintech (1 mentions)
Anti cleaved caspase 3, by ZenBio (1 mentions)
Anti p62, by Proteintech (1 mentions)
Anti p akt ser473, by Cell Signaling Technology (1 mentions)
Anti caspase 3, by Cell Signaling Technology (1 mentions)
Anti β actin, by Proteintech (1 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
Anti bax, by Proteintech (1 mentions)
3 protocols using anti mtor
1
Protein Extraction and Western Blot Analysis
2
Western Blotting Analysis of Protein Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed using a dioctanoic acid protein detection kit (Cat #PC0020; Solarbio). Samples (40 to 60 µg/10 µL) were separated using 8% or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Polyvinylidene difluoride membranes were subsequently used for protein transfer, and samples were incubated overnight at 4 °C with the following primary antibodies: anti-p-Akt (Ser473) (1:2000; RRID: AB_2315049; Cell Signaling Technology), anti-Akt (1:1000; RRID: AB_915783; Cell Signaling Technology), anti-p-mTOR (1:2000; Cat #R25033; ZenBio), anti-mTOR (1:2000; Cat #380,411; ZenBio), anti-p62 (1:2000; RRID: AB_10694431; Proteintech), anti-LC3 (1:1000; RRID: AB_2137737; Proteintech), anti-Bax (1:10 000; RRID: AB_2061561; Proteintech, Planegg-Martinsried Germany), anti-Bcl-2 (1:1000, Proteintech, RRID: AB_2227948), anti-cleaved caspase 3 (1:1000; Cat #341,034; ZenBio), anti-caspase 3 (1:1000; RRID: AB_331439; Cell Signaling Technology), anti-caspase 9 (1:500; RRID: AB_2068632; Proteintech), and anti-β-actin (1:5000; Cat #CL594-66009; Proteintech). Goat anti-mouse or anti-rabbit IgG secondary antibodies were incubated with horseradish peroxide (1:500; RRID: AB_449890; Abcam, Cambridge, UK). Signals were detected using a bioimaging system (ChemiDoc XRS+; Bio-Rad), and ImageJ software was used to analyze the band intensity.
3
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total proteins were extracted from different treatment groups using lysis buffer (Solarbio) and centrifuged at 12,000 rpm for 30 min at 4°C. The protein concentration was measured using a BCA protein assay kit (BestBio, Shanghai, China). Additionally, the protein was separated by 10% SDS-PAGE and transferred to a PVDF membrane (Millipore Corporation, Billerica, MA). The membrane was blocked with closed solution (Beyotime, Shanghai, China) and then incubated with different primary antibodies, respectively. The following primary antibodies were used: anti-MYOG (Biorbyt, Cambridge, UK; 1:1,000) and anti-MYOD1 (Santa Cruz Biotechnology, Dallas, TX, 1:1,000), anti-MTOR, anti-p-MTOR, anti-P70S6K, anti-P-P70S6K, and anti-β-tubulin (Zenbio, Chengdu, China, 1:1,000) for 12 h. The secondary antibodies used were: mouse anti-rabbit horseradish peroxidase (HRP ), goat anti-mouse HRP (Zenbio, 1:1,000). Finally, protein stripes signals were promoted by the ECL kit (Beyotime) and detected by the ImageJ (NIH Image J System, Bethesda, MD). β-Tubulin was used as the internal 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!