Ab154812

Manufactured by Abcam

Sourced in United Kingdom, United States

Ab154812 is a laboratory equipment product. It is a high-quality instrument designed for scientific research applications. The core function of this product is to facilitate specific tasks within a laboratory environment.

Automatically generated - may contain errors

Lab products found in correlation

Ab9485, by Abcam (2 mentions) Fluorchem hd2, by Bio-Techne (1 mentions) Protease and phosphatase inhibitors cocktail, by Thermo Fisher Scientific (1 mentions) Protease inhibitors cocktail, by Roche (1 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Anti creb, by Cell Signaling Technology (1 mentions) A0760 40, by US Biological (1 mentions) Ab6721, by Abcam (1 mentions) Ripa buffer, by Thermo Fisher Scientific (1 mentions) Ab68153, by Abcam (1 mentions) A2066, by Merck Group (1 mentions) Alexafluor 680 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Phosstop, by Roche (1 mentions) Protease inhibitory cocktail, by Roche (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) 4336 bpc 100, by Bio-Techne (1 mentions) Ab13847, by Abcam (1 mentions) Ab53154, by Abcam (1 mentions) Ab38449, by Abcam (1 mentions) Ab134903, by Abcam (1 mentions)

6 protocols using ab154812

Protein Purification and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Proteins were purified from 3t3 cells and BLA tissue. Cells and tissue were lysed using a lysis buffer containing Tris pH 7.4 50 mM, NaCl 150 mM, EDTA 5 mM, Triton X-100 1%, protease inhibitors cocktail (Roche, Mannheim, Germany) and protease and phosphatase inhibitors cocktail (ThermoScientific, Rockford, USA)). Cell/tissue homogenate was sonicated and total protein content was quantified using a commercial kit (Pierce^TM BCA Protein Assay Kit, Thermoscientific) with assistance of the software GEN5 (BioTek, Vermont, USA).
Western Blot analysis was conducted using the following primary antibodies and dilutions: anti-PTEN: 1:1000, ab154812 (Abcam, Cambridge, UK), anti-beta actin 1:2000, A0760–40 (USBiological, Massachusetts, USA) and anti-GAPDH 1:3000, ab9485 (Abcam), anti-CREB 1:250, 48H2 (Cell Signalling, Boston, Massachussets). Secondary antibody signals were detected using the Clarity^TM Western ECL detection kit (BioRad, California, USA) and the software Fluorchem^TM HD2 (Alpha Innotech, Biozym, Vienna, Austria). Quantification of optical densities was performed using the program ImageJ (Maryland, USA) and values were normalized to those of housekeeping genes (beta actin and GAPDH).

Ronovsky M., Zambon A., Cicvaric A., Boehm V., Hoesel B., Moser B.A., Yang J., Schmid J.A., Haubensak W.E., Monje F.J, & Pollak D.D. (2019). A role for miR-132 in learned safety. Scientific Reports, 9, 528.

+ Open protocol

+ Expand

Western Blot Analysis of PTEN, Stat3 in RLS40 and B16 Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates from RLS₄₀ and B16 cells were collected in radioimmunoprecipitation assay (RIPA) buffer (Thermo Scientific, Waltham, MA, USA), separated in 12.5% sodium dodecyl sulfate (SDS)/PAGE and transferred to a poly(vinylidene difluoride) (PVDF) membrane using a semidry transfer. Western Blot analysis was performed as described earlier [30 (link),39 (link)] using primary antibodies against PTEN (ab154812, 1:800; Abcam, Cambridge, UK), Stat3 (ab68153, 1:500; Abcam, Cambridge, UK), and GAPDH (ab9485, 1:1000; Abcam, Cambridge, UK), and secondary HRP-conjugated goat anti-rabbit antibodies (ab6721; Abcam, Cambridge, UK).

Gaponova S., Patutina O., Sen’kova A., Burakova E., Savin I., Markov A., Shmendel E., Maslov M., Stetsenko D., Vlassov V, & Zenkova M. (2022). Single Shot vs. Cocktail: A Comparison of Mono- and Combinative Application of miRNA-Targeted Mesyl Oligonucleotides for Efficient Antitumor Therapy. Cancers, 14(18), 4396.

+ Open protocol

+ Expand

Quantitative Western Blot Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells remaining after the seeding for clonogenic assays were pelleted by centrifugation, lysed in 20 mM Tris HCl pH 7.5, 400 mM NaCl, 5 mM DTT, 20 % glycerol, 0.1 % NP40, 1 mM Pefabloc, Protease Inhibitory Cocktail (Roche), phosSTOP (Roche), 100 nM Ku-0058948, 1 µM ADP-HPD (Trevigen) and analysed by western blot as previously described [11 (link)]. Antibodies used were rabbit anti-PAR (1/1000, 4336-BPC-100, Trevigen), anti-PARG (1/2000, [8 (link)]), anti-actin (1/500, A2066, Sigma), anti-PTEN (1/2000, ab154812, Abcam) and anti-BRCA1 (1/5000, 07-434, Millipore) antibodies. Secondary antibodies were either an Alexa Fluor 680 goat anti-rabbit (1/30,000, Invitrogen) or a peroxidase-coupled goat anti rabbit (1/50,000, Invitrogen), revealed either with Odyssey Infrared Imaging System (Li-Cor, Bioscience) or by chemiluminescence and autoradiography.

Noll A., Illuzzi G., Amé J.C., Dantzer F, & Schreiber V. (2016). PARG deficiency is neither synthetic lethal with BRCA1 nor PTEN deficiency. Cancer Cell International, 16, 53.

+ Open protocol

+ Expand

Protein Expression Analysis in Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

The total protein of tissues was extracted using radio-immunoprecipitation assay lysate (strong) (R0010, Solarbio). The protein concentration was determined using the BCA kit (20201ES76, Yeasen Company, Shanghai, China). The protein was separated on polyacrylamide gel electrophoresis and transferred onto the polyvinylidene difluoride membrane. After that, the membrane was blocked with 5% bovine serum albumin for 1 h and incubated with the following diluted primary antibodies at 4 °C overnight: PTEN (1:5000, ab154812, Abcam), Cleaved caspase-3 (1:500, ab13847, Abcam), Bax (1:500, ab53154, Abcam), Bcl-2 (1:1000, ab196495, Abcam), AKT (1:500, ab38449, Abcam), pAKT (1:1000, #9271, Cell Signaling Technology, Beverly, MA, USA), mTOR (1:10,000, ab134903, Abcam), pmTOR (1:1000, #2971, Cell Signaling Technology), and β-actin (1:5000, ab8226, Abcam). Following tris-buffered saline-tween buffer washing (5 min × 3 times), the membrane was added with horseradish peroxidase-labeled goat anti-rabbit IgG (ab205718, 1:20,000, Abcam). The membrane was added with enhanced chemiluminescence for development. ImageJ 1.48 u software (National Institutes of Health) was used for protein quantitative analysis, and the grayscale ratio of each protein to internal reference β-actin was used for protein quantitative analysis. The experiment was repeated three times.

Xiao X., Li W., Rong D., Xu Z., Zhang Z., Ye H., Xie L., Wu Y., Zhang Y, & Wang X. (2021). Human umbilical cord mesenchymal stem cells-derived extracellular vesicles facilitate the repair of spinal cord injury via the miR-29b-3p/PTEN/Akt/mTOR axis. Cell Death Discovery, 7, 212.

+ Open protocol

+ Expand

Western Blot Protocol for PTEN Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in RIPA buffer containing protease inhibitors (Sigma, Oakville, Canada). Equal amounts of proteins were resolved on 10% SDS-PAGE and transferred to a nitrocellulose membrane (BioRad, CA, USA). Membranes were blocked in TBS containing 5% non-fat dry milk and exposed overnight at 4 °C to rabbit-anti-PTEN (ab154812, Abcam, MA, USA) or mouse-anti-β-Actin (A5316, Sigma, Oakville, Canada) antibodies. Membranes were washed in TBST (TBS-0.05% Tween-20) and incubated with either anti-rabbit or anti-mouse peroxidase-conjugated secondary antibody for 1 hour at room temperature and the blots were developed using Immobilon Western HRP Substrate (Millipore, Etobicoke, Canada).

Abdouh M., Gao Z.H., Arena V., Arena M., Burnier M.N, & Arena G.O. (2019). Oncosuppressor-Mutated Cells as a Liquid Biopsy Test for Cancer-Screening. Scientific Reports, 9, 2384.

+ Open protocol

+ Expand

Retinal PTEN/Akt/Bcl-2 Immunohistochemistry

Check if the same lab product or an alternative is used in the 5 most similar protocols

Retinal sections (10 lm) through the optic nerve head were selected for each eye. Deparaffinized retinal sections were immersed in proteinase K solution for antigen retrieval and blocked in 10% goat serum for 1 hour. The sections then were incubated with the following primary antibodies at 48C overnight: PTEN (1:100, ab154812; Abcam, Cambridge, UK), p-Akt (1:200, 4060S; Cell Signaling Technology, Beverly, MA, USA) and B-cell lymphoma 2 (Bcl-2, 1:100, ab7973; Abcam). After thorough washing with PBS, the sections were exposed to their corresponding secondary Cy3 antibodies (1:300) for 1 hour at room temperature, after which they were counterstained with DAPI, washed again, coverslipped, and examined under a fluorescence microscope.

Mao D, & Sun X. (2015). Reactivation of the PI3K/Akt Signaling Pathway by the Bisperoxovanadium Compound bpV(pic) Attenuates Photoreceptor Apoptosis in Experimental Retinal Detachment. Investigative ophthalmology & visual science, 56(9).

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!