Tyrosine Phosphorylation ProArray was bought from Full Moon Biosystems (Sunnyvale, CA, USA) (#PST228). To retrieve enough protein for the array, ten E12.5 control and twelve E12.5 mutant lungs were pooled together. Protein extract was quantified with using DC protein assay (Bio-Rad Laboratories): 40 μg of protein lysate was conjugated with biotin and incubated on the phospho-protein array. The fluorescence intensity of each spot was quantified using Image-J and spots were compared after subtracting background and negative control signals. The signal intensity for the six replicates were averaged and showed as average ± standard deviation.
Tyrosine phosphorylation proarray
Manufactured by Full Moon BioSystems
Sourced in United States
The Tyrosine Phosphorylation ProArray is a lab equipment product designed to detect and quantify tyrosine phosphorylation levels in protein samples. It utilizes an array-based approach to provide a comprehensive analysis of tyrosine phosphorylation patterns.
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3 protocols using tyrosine phosphorylation proarray
1
Tyrosine Phosphorylation Profiling in Lung Development
2
Profiling Tyrosine Kinase Signaling
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For profiling the phosphorylation activity of p190 and p210 isoforms in cell line models, a Tyrosine Phosphorylation Proarray (Full Moon Biosystems, #PST228) featuring 228 phospho-tyrosine sites was used. Sample preparation and processing were performed according to manufacturer’ instructions. Array scanning and image analysis were performed by Full Moon Bioscience and data were normalized to the median antibody signaling value. Western blotting was used to validate findings in cell lines and CML patient samples (three p190 and three p210). For Ph+ALL samples, phosphorylation of selected proteins was investigated by flowcytometry using iQue Screener Plus flow cytometer (Intellicyt). Further details are described in Supplementary materials.
3
Kinase and Tyrosine Phosphorylation Profiling
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Kinase Antibody Array Kit (AVK276, including 276 kinase antibodies) and Tyrosine Phosphorylation ProArray (PST228, including 228 site-specific tyrosine phosphorylation antibodies) were obtained from Full Moon Biosystems (Sunnyvale, CA USA)28 (link). Protein microarray analysis was carried out by Wayen Biotechnologies (Shanghai, China). Briefly, proteins from the heart tissues of control or type 1 diabetic mice were extracted and labeled with Biotin. The samples were diluted 1:20 in coupling solution before being applied to the array for conjugation. Arrays were washed three times, and Cy3-Streptavidin was used for detection. Slides were scanned using the GenePix 4000B Array Scanner (Axon Instruments, Foster City, CA, USA). The analyzed results were expressed by the ratio of phosphorylated/total protein.
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