phospho her2  (Cell Signaling Technology Inc)

Journal: Oncotarget

Article Title: Development of parallel reaction monitoring (PRM)-based quantitative proteomics applied to HER2-Positive breast cancer

doi: 10.18632/oncotarget.26031

Figure Lengend Snippet: Proteotypic peptides of EGFR, HER2, HER3, PTEN and phospho-HER2, their m/z at defined charge, the range of light and heavy peptides injected for calibration curve

Article Snippet: Antibodies used were anti HER2 (2165), Phospho-HER2 (9923), EGFR (2232), and PTEN (9583) from Cell Signaling ® with 1:1000 dilution.

Techniques: Injection

Journal: Oncotarget

Article Title: Development of parallel reaction monitoring (PRM)-based quantitative proteomics applied to HER2-Positive breast cancer

doi: 10.18632/oncotarget.26031

Figure Lengend Snippet: ( A ) Analysis of 17 BCLs. Horizontal axis represents the HER2/actin ratio obtained with western blot. Vertical axis is the light/heavy ratio of the GLQSLPTHDPSPLQR HER2 peptide of the same BCL. We also represented ICC classification of these BCL (light grey: zero expression of HER2, dark grey: equivocal in ICC; black: overexpression of HER2 in ICC. ( B ) Analysis of 17 BCLs. Horizontal axis represents the HER2/actin ratio obtained with western blot. Vertical axis is the light/heavy ratio of the mean of pooled HER2 peptides (GIWIPDGENVK; SGGGDLTLGLEPSEEEAPR; GTPTAENPEYLGLDVPV, GLQSLPTHDPSPLQR) of the same BCL. We also represented ICC classification of these BCLs (light grey: zero expression of HER2, dark grey: equivocal in ICC; black: overexpression of HER2 in ICC). ( C ) Box plot representing the light/heavy ratio of GLQSLPTHDPSPLQR peptide depending on ICC status. Significant correlation between each condition was observed: none expressors, equivocal status or overexpressors. ( D ) Box plot representing the light/heavy ratio of pooled HER2 peptides (GIWIPDGENVK; SGGGDLTLGLEPSEEEAPR; GTPTAENPEYLGLDVPV, GLQSLPTHDPSPLQR) depending on ICC status. Significant correlation between each condition: ns = non significant; * p < 0.05; ** p < 0.01; *** p < 0.001. Data are represented as mean of six technical replicates ± standard deviation.

Article Snippet: Antibodies used were anti HER2 (2165), Phospho-HER2 (9923), EGFR (2232), and PTEN (9583) from Cell Signaling ® with 1:1000 dilution.

Techniques: Western Blot, Expressing, Over Expression, Standard Deviation

Journal: Oncotarget

Article Title: Development of parallel reaction monitoring (PRM)-based quantitative proteomics applied to HER2-Positive breast cancer

doi: 10.18632/oncotarget.26031

Figure Lengend Snippet: Box Plot representing the large magnitude of range of HER2 expression, from under LLOQ to 9860 amol/µg. Data are represented as mean of six technical replicates ± SEM.

Article Snippet: Antibodies used were anti HER2 (2165), Phospho-HER2 (9923), EGFR (2232), and PTEN (9583) from Cell Signaling ® with 1:1000 dilution.

Techniques: Expressing

Journal: Oncotarget

Article Title: Development of parallel reaction monitoring (PRM)-based quantitative proteomics applied to HER2-Positive breast cancer

doi: 10.18632/oncotarget.26031

Figure Lengend Snippet: ( A ) represents the mean of the pooled non-phosphorylated peptides (GIWIPDGENVK; SGGGDLTLGLEPSEEEAPR; GTPTAENPEYLGLDVPV, GLQSLPTHDPSPLQR); ( B ) the phosphorylated HER2 peptide GTPTAENPEpY(1248)LGLDVPV; ( C ) the mean of GTPTAENPEpY(1248)LGLDVPV/GTPTAENPEYLGLDVPV ratio; ( D ) the mean of GTPTAENPEpY(1248)LGLDVPV/GTPTAENPEYLGLDVPV ratio under trastuzumab compared to control condition in sensitive and resistant BCL, determined by Ginestier et al. ; ( E ) western blots of HER2 and phospho-HER2 peptide GTPTAENPEpY(1248)LGLDVPV on SKBR3 breast cell line; ( F ) the mean of the pooled non-phosphorylated peptides (GIWIPDGENVK; SGGGDLTLGLEPSEEEAPR; GTPTAENPEYLGLDVPV, GLQSLPTHDPSPLQR) and phospho-HER2 peptide GTPTAENPEpY(1248)LGLDVPV on SKBR3 breast cell line. Significant correlation between each condition: ns = non significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. EC = Effective Concentration. PRM data are represented as mean of six technical replicates ± SEM.

Article Snippet: Antibodies used were anti HER2 (2165), Phospho-HER2 (9923), EGFR (2232), and PTEN (9583) from Cell Signaling ® with 1:1000 dilution.

Techniques: Control, Western Blot, Concentration Assay

Journal: Oncotarget

Article Title: Development of parallel reaction monitoring (PRM)-based quantitative proteomics applied to HER2-Positive breast cancer

doi: 10.18632/oncotarget.26031

Figure Lengend Snippet: ( A ) Light/heavy ratio of the 8 PDX. Each image represents the score corresponding HER2 expression on IHC. * Sample 6 was classified 3+ on IHC of PDX as it was 2+ on IHC of corresponding patient. ( B ) HER2 absolute quantification of the 46 tumor samples. The table represents the corresponding characteristics of the tumors: HER2 defined with IHC: N = negative; P = positive; ND = not determined (0 = 0 cross negative status; 2 = 2+ equivocal status; 3 = 3+ positive status) and/or with FISH. LLOQ line represents the limit of absolute quantification. Data are represented as mean of four replicates ± standard deviation.

Article Snippet: Antibodies used were anti HER2 (2165), Phospho-HER2 (9923), EGFR (2232), and PTEN (9583) from Cell Signaling ® with 1:1000 dilution.

Techniques: Expressing, Quantitative Proteomics, Standard Deviation

Journal: Scientific Reports

Article Title: A multifunctional DNA nano-scorpion for highly efficient targeted delivery of mRNA therapeutics

doi: 10.1038/s41598-018-28542-3

Figure Lengend Snippet: Characterization of AptDzy-DNS induced translocation. Confocal microscopy images of SK-BR-3 cells after incubation with 0.5 μM Dzy-DNS without aptamer, HAptDzy-DNS, NAptDzy-DNS, and the mixture of HAptDzy-DNS and NApt-Dzy-DNS for 2 h. Scale bars: 20 μm. (Note: NApt, anti-nucleolin aptamer; HApt, anti-HER2 aptamer; Apt, aptamer).

Article Snippet: And then the membrane was blocked with 5% fat-free milk and incubated with primary antibodies (diluted at ratios of 1:1,000 (β-Actin, bioworld #AP0060; Phospho-HER2/ErbB2 Antibody Sampler Kit, CST #9923)) at 4 °C overnight or 25 °C for 2 h. The complex of antigen-antibody was visualized with New Super ECL Assay and detected by an ECLion system.

Techniques: Translocation Assay, Confocal Microscopy, Incubation

Journal: Scientific Reports

Article Title: A multifunctional DNA nano-scorpion for highly efficient targeted delivery of mRNA therapeutics

doi: 10.1038/s41598-018-28542-3

Figure Lengend Snippet: Cell cytotoxicity assay and proliferation assay. ( A ) CCK-8 assay of cell viability for SK-BR-3 cells incubated with DNS at various concentration. ( B ) CCK-8 assay of cell viability for SK-BR-3 cells incubated with AptDzy-DNS at different hours. ( C ) RT-PCR characterization of HER2 mRNA expression for SK-BR-3 cells incubated without and with AptDzy-DNS. The data error bars indicate means ± SD (n = 5). * P < 0.05, ** P < 0.01 (two-tailed Student’s t-test), ( D ) Western blot characterization of HER2 expression for SK-BR-3 cells incubated with 0.25 (Lane 1), 0 (Lane 2), 0.5 (Lane 3), 0.25 (Lane 4), and 0.125 (Lane 5) μM AptDzy-DNS.

Article Snippet: And then the membrane was blocked with 5% fat-free milk and incubated with primary antibodies (diluted at ratios of 1:1,000 (β-Actin, bioworld #AP0060; Phospho-HER2/ErbB2 Antibody Sampler Kit, CST #9923)) at 4 °C overnight or 25 °C for 2 h. The complex of antigen-antibody was visualized with New Super ECL Assay and detected by an ECLion system.

Techniques: Cytotoxicity Assay, Proliferation Assay, CCK-8 Assay, Incubation, Concentration Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Two Tailed Test, Western Blot