Phospho-array detection was performed in collaboration with Wayen Biotechnology (Shanghai, China). Proteins were extracted from RD cells, labeled with biotin, and hybridized to the Phosphorylation ProArray (Full Moon BioSystems, Sunnyvale, CA, USA) using an Antibody Array Kit (Full Moon BioSystems) to determine the specific signaling phospho-antibody profiles. Finally, we scanned the corresponding fluorescence intensity using a GenePix 4000B instrument (Axon Instruments, Houston, TX, USA) using GenePix Pro 6.0 software to acquire phosphorylation levels for specific proteins. In our study, Ratio ≥1.15 was the threshold for upregulation and Ratio ≤1/1.15 was the threshold for downregulation.
Phosphorylation proarray
Manufactured by Full Moon BioSystems
Sourced in United States
The Phosphorylation ProArray is a laboratory equipment product that facilitates the analysis and detection of protein phosphorylation. It provides a platform for comprehensive, high-throughput screening of phosphorylation events across multiple protein targets.
Automatically generated - may contain errors
5 protocols using phosphorylation proarray
1
Phosphorylation Profiling of RD Cells
2
Phosphorylation Profiling of Cancer Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phospho-array detection was performed in cooperation with Wayen Biotechnology (Shanghai, China). Cells transfected with RalA siRNA or miR-181a were collected 72 h post-transfection for the extraction of protein; 100 μg cell lysate was labeled with biotin reagent and hybridized on Phosphorylation ProArray (Full Moon BioSystems, CA, USA) using an Antibody Array Kit (Full Moon BioSystems) for the detection of 248-site-specific Cancer Signaling Phospho Antibody profiles. Finally, fluorescence intensity was scanned with a GenePix 4000B (Axon Instruments, Houston, TX, USA) using GenePix Pro 6.0. The raw data were treated via Grubbs' method. The phosphorylation ratio was calculated as follows: phosphorylation ratio = phospho value/unphospho value.
3
Phospho-array analysis of cancer signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phospho-array detection was performed in cooperation with Wayen Biotechnology (Shanghai, China). At 48 h post-transfection, all treated cells were collected for protein extraction. Protein samples of 50 mg each were tagged with biotin reagent and hybridized on a Phosphorylation ProArray (Full Moon BioSystems, USA) using an Antibody Array Kit (FullMoon BioSystems, USA) for the detection of 248 site-specific cancer signaling phospho-antibody profiles. Finally, fluorescence intensity was scanned by GenePix 4000B (Axon Instruments, Houston, USA) using GenePix Pro 6.0. The raw data were manipulated using Grubbs’ method. The phosphorylation ratio was calculated as follows: phosphorylation ratio ¼ phospho value/unphospho value.
4
Phospho-Array Analysis of PPFIA1 and miR-181a
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phospho-array detection was performed in collaboration with Wayen Biotechnology (Shanghai, China). After transfection with PPFIA1 siRNA or miR-181a mimic (72 h), treated K562 cells were collected for protein extraction. Protein samples (50 mg each) were labeled with biotin and hybridized to the Phosphorylation ProArray (Full Moon BioSystems, USA) using an Antibody Array Kit (Full Moon BioSystems, CA, USA) for the detection of 248 site-specific cancer signaling phospho-antibody profiles. Finally, fluorescence intensity was scanned with a GenePix 4000B (Axon Instruments, Houston, TX, USA) using GenePix Pro 6.0. Raw data were manipulated using Grubbs’ method. The reduction phosphorylation ratio was calculated as follows: reduction phosphorylation ratio = phospho value/non-phosphorylated value.
5
Phosphoproteomic Analysis of Granulosa Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Granular cells of follicles with or without progesterone treatment on culture day 10 were collected for protein extraction. Phospho-array detection and data analysis were performed in collaboration with Wayen Biotechnology (Shanghai, China) using the system they set up [23] (link). Briefly, protein samples were labeled with biotin and hybridized to the Phosphorylation ProArray (Full Moon BioSystems, USA) using an Antibody Array Kit (Full Moon BioSystems, CA, USA). The antibody array was composed of 1318 antibodies and most of the antibodies were used to detect the phosphorylated form of the proteins and their unphosphorylated counterparts. Fluorescence intensity was scanned with a GenePix 4000B (Axon Instruments, Houston, TX, USA) using GenePix Pro 6.0. Raw data of were manipulated using Grubbs’ method to exclude outliers. The phosphorylation ratio was calculated as follows: phosphorylation ratio = phospho value/nonphosphorylated value. Phosphoproteins that were upregulated or down regulated by more than 20% (P < 0.05) were included in the analysis. The key signaling pathways were further analyzed using the Kyoto Encyclopedia of Genes and Genomes (KEGG).
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!