His-tag recombinant human kallistatin was expressed in Pichia pastoris strain GS115 and purified by a series of chromatographic steps, mainly Phenyl Superose and Heparin Sepharose FF chromatography [24 (link), 25 ]. Rabbit anti-human integrin β3, rabbit anti-human phospho-Integrin β3, and mouse anti-human integrin β3 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit anti-human AKT, rabbit anti-human phospho-AKT, rabbit anti-human Erk1/2, rabbit anti-human phospho-Erk1/2, rabbit anti-human FAK, rabbit anti-human phospho-FAK, rabbit anti-human Src and rabbit anti-human phospho-Src were obtained from Cell Signaling Technology (Boston, MA). 3-(4,5-dimethylthiazol-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide was purchased from Sigma. Click-iT EdU Alexa Fluor 594 Imaging Kit (including Hoechst 33,342 and Apollo reaction cocktail) and Lipofectamine™ 2000 were from Life Technologies (Gaithersburg, MD). PVDF membranes and the Immobilon ECL detection system were purchased from Millipore Co (Billerica, MA). Integrin β3 and negative control siRNA were purchased from GenePharma (Shanghai, China).
Rabbit anti human akt
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit anti-human AKT is an antibody that specifically recognizes the AKT protein in human cells. AKT is a serine/threonine protein kinase that plays a key role in various cellular processes, including cell growth, proliferation, and survival.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti human phospho akt, by Cell Signaling Technology (2 mentions)
Rabbit anti human erk1 2, by Cell Signaling Technology (2 mentions)
Rabbit anti human phospho erk1 2, by Cell Signaling Technology (2 mentions)
Rabbit anti human fak, by Cell Signaling Technology (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Rhil 6, by R&D Systems (1 mentions)
Rabbit anti human vegf, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti human stat3, by Cell Signaling Technology (1 mentions)
Mouse anti gapdh, by Merck Group (1 mentions)
Miniprotein 3 system, by Bio-Rad (1 mentions)
Polyvinylidene fluoride membrane, by GE Healthcare (1 mentions)
Alliance q9 advanced, by Uvitec (1 mentions)
Electrochemiluminescence kit, by Proteintech (1 mentions)
Caspase 3 p17, by Proteintech (1 mentions)
Unbs5162, by MedChemExpress (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
P mtor, by Cell Signaling Technology (1 mentions)
Gapdh, by Proteintech (1 mentions)
Bcl2 associated x (bax), by Proteintech (1 mentions)
5 protocols using rabbit anti human akt
1
Purification and Analysis of Recombinant Kallistatin
2
IL-6 Signaling Pathway Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
UM-HMC cell lines were plated, serum starved overnight, treated with tocilizumab or Stattic (STAT3 inhibitor V, Calbiochem, San Diego, CA, USA) at the indicated concentrations, and exposed to 20 ng/ml rhIL-6 (R&D Systems, Minneapolis, MN, USA). NP-40 lysis buffer was used to prepare whole cell lysates that were resolved using PAGE. Membranes were incubated with the following primary antibodies for 1 hour at room temperature or overnight at 4°C: mouse anti-human phospho-STAT3, rabbit anti-human STAT3, rabbit anti-human phospho-ERK1/2, rabbit anti-human ERK1/2, rabbit anti-human phospho-AKT, rabbit anti-human AKT, rabbit anti-human Bcl-xL, rabbit anti-human gp130 (Cell Signaling, Beverly, MA, USA), rabbit anti-human IL-6Rα, rabbit anti-human VEGF (Santa Cruz Biotechnology, Santa Cruz, CA, USA), hamster anti-human Bcl-2 (BD Pharmingen, Franklin Lakes, NJ, USA); or mouse anti-GAPDH (Chemicon, Billerica, MA, USA).
3
Western Blot Analysis of CD279, pAKT, AKT
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein was extracted by using RIPA lysis buffer. The equal amount of protein samples was analyzed on 10% SDS-PAGE by mini-Protein III system (Bio-Rad, USA). The proteins were transferred to polyvinylidene fluoride membrane (GE Healthcare, USA). The membrane was probed with mouse anti-human CD279 antibody (BioRad), rabbit anti-human pAKT (Thr308) (Cell Signaling Technology), rabbit anti-human AKT (Cell Signaling Technology) and rabbit anti-human GAPDH antibody (BioRad). The signals were detected in different exposure times depending on each protein by horseradish peroxide (HRP) conjugated secondary antibodies and enhanced chemiluminescence substrate with Alliance Q9 Advanced (UVITEC, UK) or X-ray film. All original unmanipulated Western blot images were provided as the Supplementary Information.
4
Signaling Pathways in UNBS5162 Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
UNBS5162 was obtained from MedChemExpress (Princeton, NJ, USA). All experiments were performed using 10 μM UNBS5162 as the work concentration. Reference antibodies were obtained: rabbit antihuman Akt, p-Akt, mTOR, p-mTOR (Cell Signaling Technology, Danvers, MA, USA), rabbit antihuman p70, Bcl2, Bax, caspase 3 p17, mouse antihuman cyclin D1, and GAPDH (ProteinTech Wuhan, China). The electrochemiluminescence kit was also purchased from ProteinTech.
5
Flaccidoxide-13-acetate Signaling Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Flaccidoxide-13-acetate was supplied by Dr. Jui-Hsin Su. Rabbit anti-human Akt, FAK, MMP-2, MMP-9, MMP-13, mTOR, PI3K, TIMP-1, TIMP-2, uPA and phosphorylated Akt, mTOR, and PI3K were purchased from Cell Signaling Technology (Danvers, MA, USA). E-cadherin, N-cadherin, lamin A2, and Snail antibodies were supplied by Epitomics (Burlingame, CA, USA). HRP-conjugated goat anti-rabbit immunoglobulin (IgG) was obtained from Millipore (Bellerica, MA, USA). Thiazolyl blue tetrazolium bromide, streptomycin, penicillin, and DMSO were purchased from Sigma (St. Louis, MO, USA). Goat anti-rabbit and horseradish peroxidase-conjugated immunoglobulin IgG and polyvinylidene difluoride (PVDF) membranes were obtained from Millipore (Bellerica, MA, USA). Fetal bovine serum and DMEM were purchased from Biowest (Nuaillé, France).
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!