The rat livers and AML12 cells were lysed by RIPA buffer, separated by 10% polyacrylamide gel electrophoresis and then transferred to polyvinylidene fluoride (PVDF) membrane. After blocking in 5% Bovine Serum Albumin (BSA), the membrane was incubated with the phospho-JAK2 (Tyr931) (1:1,000, affinity), total JAK2 (1:1,000, proteintech), phospho-STAT3 (Tyr705) (1:1,500, abcam), total STAT3 (1:2000, abcam), IL-6 (1:500, bioworld), IL-18 (1:2000, bioworld), IL-1β (1:500, sino biological), TNFα (1:1,000, sino biological), HIF-1a (1:1,000, Novusbio), HIF-2a (1:1,000, Cell Signaling Technology), VEGFa (1:1,000, Novusbio) primary antibody seperately overnight at 4°C, and then was incubated with HRP conjugated secondary antibody for 1 h at room temperature. Western blots were displayed by advanced enhanced chemiluminescence kit and then semi-quantified by Clinx (Shanghai Qinxiang Scientific Instrument Co., Ltd).
Hif2a
Manufactured by Cell Signaling Technology
HIF2A is a protein that plays a central role in the cellular response to hypoxia. It functions as a transcriptional activator that regulates the expression of genes involved in processes such as angiogenesis, erythropoiesis, and metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Bcl 2, by Cell Signaling Technology (2 mentions)
Cleaved caspase 3, by Cell Signaling Technology (2 mentions)
E cadherin, by Cell Signaling Technology (2 mentions)
Il 1β, by Sino Biological (1 mentions)
Phospho jak2, by Proteintech (1 mentions)
Tnf α, by Sino Biological (1 mentions)
Vegfa, by Novus Biologicals (1 mentions)
Phospho stat3 tyr705, by Abcam (1 mentions)
Total stat3, by Abcam (1 mentions)
Interleukin 6 (il 6), by Bioworld Technology (1 mentions)
Hif 1a, by Novus Biologicals (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Mmp 9, by Cell Signaling Technology (1 mentions)
3 protocols using hif2a
1
Signaling Pathways in Liver and AML12 Cells
2
Protein Expression Analysis via Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Forward Reverse were subjected to SDS-polyacrylamide gel electrophoresis, transferred onto a polyvinylidene fluoride membrane, and incubated first with rabbit antibodies against E-cadherin, Bcl2, cleaved caspase-3, HIF2A, p62, LC3, and actin (all from Cell Signaling Technology) overnight at 4 °C, followed by another 2 h incubation with a secondary antibody (1:5000). GAPDH served as a loading control. The secondary antibodies were horseradish peroxidase (HRP)-conjugated anti-rabbit (Jackson ImmunoResearch Labs, West Grove, PA, USA). Densitometric analysis of the western blots was performed with NIH Image J software (Bethesda, MD, USA). The protein levels were first normalized to actin and subsequently to the experimental controls.
3
Cell Culture and Reagents for Renal Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 786-O and ACHN cell lines were obtained from the American Type Culture Collection (Rockville, MD, USA). Line 786-O was grown in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) in a humidified chamber with 5% CO 2 at 37 °C; ACHN cells were grown in MEM supplemented with 10% FBS. FBS was purchased from Gibco (Life Technologies, CA, USA). The penicillin-streptomycin solution was Hyclone (Thermo Fisher Scientific, Massachusetts, USA). The cell strain was purchased from Corning (Corning, NY, USA). Cyanidin was purchased from Sigma-Aldrich Corporation (St. Louis, MO, USA). Antibodies against MMP-9, Cyclin D1, E-cadherin, HIF2A, Bcl2, cleaved caspase-3, LC3, and p62 were purchased from Cell Signaling Technology (Danvers, MA).
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!