Human proteome profiling array system

Manufactured by R&D Systems

The human proteome profiling array system is a tool designed for the comprehensive analysis of protein expression profiles in human samples. It enables the simultaneous detection and quantification of a large number of human proteins in a single experiment. The core function of this system is to provide researchers with a detailed snapshot of the protein composition and relative abundance within a given sample.

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Lab products found in correlation

Proteome profiler human phospho kinase array kit, by R&D Systems (2 mentions) Proteome profiler human phospho kinase array kit, by RnD Systems (1 mentions)

3 protocols using human proteome profiling array system

Profiling Human Kinase Phosphorylation

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We used a human proteome profiling array system (R&D Systems) that contains duplicate validated positive and negative controls and captures antibodies that can simultaneously detect the phosphorylation state of 43 human kinases (Proteome Profiler Human Phospho-Kinase Array Kit; for coordinate annotation, see https://www.rndsystems.com/; ref. 31 ). Five million cells were plated in 10-cm dishes and grown for 48 hours. Cells were deprived of serum by culturing for 24 hours in growth media supplemented with 0.05% FBS. Whole-cell extracts were then prepared, and detection of protein phosphorylation was carried out according to the manufacturer's instructions. In brief, the array membranes were blocked, incubated with 350 µg total cellular protein per array overnight at 4°C on a rocking platform, washed, and incubated with phospho-specific detection antibodies. Captured phosphorylated proteins were detected by ECL and imaged on X-ray films. The average pixel densities of duplicate spots were measured using ImageJ software and are expressed relative to the positive control on each array.

Schram A.M., Odintsov I., Espinosa-Cotton M., Khodos I., Sisso W.J., Mattar M.S., Lui A.J., Vojnic M., Shameem S.H., Chauhan T., Torrisi J., Ford J., O'Connor M.N., Geuijen C.A., Schackmann R.C., Lammerts van Bueren J.J., Wasserman E., de Stanchina E., O'Reilly E.M., Ladanyi M., Drilon A, & Somwar R. (2022). Zenocutuzumab, a HER2xHER3 Bispecific Antibody, Is Effective Therapy for Tumors Driven by NRG1 Gene Rearrangements. Cancer Discovery, 12(5), 1233-1247.

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Profiling Kinase Phosphorylation in Cells

Cited 1 time

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We used a human proteome profiling array system (R&D Systems) that contains duplicate validated positive and negative controls and capture antibodies that can simultaneously detect the phosphorylation state of 43 human kinases (Proteome Profiler Human Phospho-kinase Array kit; for coordinate annotation see www.rndsystems.com) (31 (link)). Five million cells were plated in 10-cm dishes and grown for 48 h. Cells were deprived of serum by culturing for 24 h in growth media supplemented with 0.05% FBS. Whole-cell extracts were then prepared, and detection of protein phosphorylation was carried out according to the manufacturer’s instructions. In brief, the array membranes were blocked, incubated with 350 μg total cellular protein per array overnight at 4 °C on a rocking platform, washed, and incubated with phospho-specific detection antibodies. Captured phosphorylated proteins were detected by ECL and imaged on x-ray films. The average pixel densities of duplicate spots were measured using ImageJ software and are expressed relative to the positive control on each array.

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Phospho-kinase profiling in cell lines

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We used a human proteome profiling array system (R&D Systems) that contain duplicate validated positive and negative controls, and capture antibodies that can simultaneously detect the phosphorylation state of 43 human kinases (Proteome Profiler Human Phospho-kinase Array kit). Five million cells were plated in 10-cm dishes and grown for 48 h. Cells were deprived of serum by culturing for 24 h in growth media supplemented with 0.05% FBS, then treated with 1 μM GSK2849330 or afatinib for 0.5 h, and detection of protein phosphorylation was carried out according to the manufacturer’s instructions. In brief, the array membranes were blocked, incubated with 350 μg total cellular protein per array overnight at 4 °C on a rocking platform, washed, and incubated with phospho-specific detection antibodies. Captured phosphorylated proteins were detected by ECL and imaged on x-ray films. The average pixel densities of duplicate spots were measured using ImageJ software (http://imagej.nih.gov/ij/), and are expressed relative to the positive control on each array.

Odintsov I., Mattar M.S., Lui A.J., Offin M., Kurzatkowski C., Delasos L., Khodos I., Asher M., Daly R.M., Rekhtman N., de Stanchina E., Ganji G., Ladanyi M, & Somwar R. (2021). Novel preclinical patient-derived lung cancer models reveal inhibition of HER3 and MTOR signaling as therapeutic strategies for NRG1 fusion-positive cancers. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 16(7), 1149-1165.

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