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Betta Pharmaceuticals Co Ltd ihc staining protocol
Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Betta Pharmaceuticals Co Ltd ihc staining positive determination protocol
Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Novus Biologicals chromogenic ihc staining of formalinfixed paraffin embedded (ffpe) tissue protocol
Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Identification of patients <t>with</t> <t>RDAA</t> NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b <t>IHC</t> staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)
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Identification of patients with RDAA NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b IHC staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)

Journal: Signal Transduction and Targeted Therapy

Article Title: RNase1-driven ALK-activation is an oncogenic driver and therapeutic target in non-small cell lung cancer

doi: 10.1038/s41392-025-02206-x

Figure Lengend Snippet: Identification of patients with RDAA NSCLC. a Quantification of plasma RNase1 in NSCLC patients (N = 48) and normal individuals (N = 15). RNase1 concentration was measured by ELISA as described in Methods. p < 0.01, Student’s t -test. b IHC staining of RNase1 in human lung tumors (N = 48) and normal lung (N = 10) tissues. RNase1 expression level was calculated based on the intensity and percentage of stained cells as described in Methods. Representative images shown. c Correlation analysis between plasma RNase1 concentration and RNase1 expression level in paired tumor tissues (N = 47). R = 0.84, Pearson’s Chi-Square test. d IHC staining of RNase1 expression and ALK phosphorylation levels in human NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 specific antibodies were used for IHC staining. Representative images shown. e Diagnosis of RDAA NSCLC patients in 1173 NSCLC tissues. ALK p-Y1604, ALK p-Y1282/1283 and RNase1 were used as biomarkers to identify RDAA positive samples. f Correlation analysis between RNase1 expression and ALK phosphorylation in NSCLC tissues which used from ( e ). ALK p-Y1604 and 1282/1283 double positive means ALK phosphorylation positive (pALK + ), otherwise means ALK phosphorylation negative (pALK-)

Article Snippet: Based on previous experimental results, we defined the tumor as RDAA positive only when the IHC staining results of the above three antibodies were triple-positive (Fig. , the antibodies of RDAA, the protocol for immunohistochemical staining and positive determination of RDAA were supported by Betta Pharmaceuticals Co., Ltd, more details see Methods).

Techniques: Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Expressing, Staining, Phospho-proteomics, Biomarker Discovery