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    PD L1 IHC 22C3 pharmDx for use with Dako Omnis
    Description:
    PD L1 IHC 22C3 pharmDx is a qualitative immunohistochemical assay using Monoclonal Mouse Anti PD L1 Clone 22C3 intended for use in the detection of PD L1 protein in formalin fixed paraffin embedded FFPE non small cell lung cancer NSCLC tissue using EnVision FLEX visualization system for use on Dako Omnis PD L1 protein expression in NSCLC is determined by using Tumor Proportion Score TPS which is the percentage of viable tumor cells showing partial or complete membrane staining at any intensity The specimen should be considered to have PD L1 expression if TPS 1 PD L1 IHC 22C3 pharmDx is indicated as an aid in identifying NSCLC patients for treatment with KEYTRUDA pembrolizumab See the KEYTRUDA product label for specific clinical circumstances guiding PD L1 testing PD L1 IHC 22C3 pharmDx on Dako Omnis The assay is a modular IHC assay for 60 tests The assay has been tailored especially with EnVision FLEX visualization system on the Dako Omnis instrument The complete assay consists of the following components to be ordered separately PD L1 IHC 22C3 pharmDx GE00621 5 for Dako Omnis Monoclonal Mouse Anti PD L1 Clone 22C3 RTU Dako Omnis with Negative Control Reagent EnVision FLEX High pH Dako Omnis GV80011 2 OR EnVision FLEX Mini Kit High pH Dako Omnis GV82311 2 EnVision FLEX Target Retrieval Solution Low pH 50x GV80511 2 EnVision FLEX Mouse Linker Dako Omnis GV82111 2 EnVision FLEX DAB Enhancer Dako Omnis GC80611 2 PD L1 Control Slides T139130 2 optional The assay requires Low pH Target Retrieval Mouse Linker and a DAB Enhancer but the PD L1 Control Slides are optional For countries outside of the United States see the local KEYTRUDA product label for approved indications and expression cutoff values to guide therapy PD L1 IHC 22C3 pharmDx is subject to an exclusive trademark license to Dako Denmark A S KEYTRUDA is a registered trademark of Merck Sharp Dohme Corp a subsidiary of Merck Co Inc
    Catalog Number:
    PD-L1-IHC-22C3-PHARMDX,-FOR-USE-WITH-DAKO-OMNIS
    Price:
    None
    Category:
    Products Dako Omnis Solution For Ihc Ish Pharmdx Kit For Dako Omnis Pharmdx Kit Pd L1 Ihc 22C3 Pharmdx For Use With Dako Omnis
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    Structured Review

    Agilent technologies egfr
    Immunohistochemical staining, a ) negative control, positive staining of b ) <t>EGFR</t> (3+), c ) <t>HER2neu</t> (1+), d ) HER3 (2+) and e ) HER4 (only sporadic)
    PD L1 IHC 22C3 pharmDx is a qualitative immunohistochemical assay using Monoclonal Mouse Anti PD L1 Clone 22C3 intended for use in the detection of PD L1 protein in formalin fixed paraffin embedded FFPE non small cell lung cancer NSCLC tissue using EnVision FLEX visualization system for use on Dako Omnis PD L1 protein expression in NSCLC is determined by using Tumor Proportion Score TPS which is the percentage of viable tumor cells showing partial or complete membrane staining at any intensity The specimen should be considered to have PD L1 expression if TPS 1 PD L1 IHC 22C3 pharmDx is indicated as an aid in identifying NSCLC patients for treatment with KEYTRUDA pembrolizumab See the KEYTRUDA product label for specific clinical circumstances guiding PD L1 testing PD L1 IHC 22C3 pharmDx on Dako Omnis The assay is a modular IHC assay for 60 tests The assay has been tailored especially with EnVision FLEX visualization system on the Dako Omnis instrument The complete assay consists of the following components to be ordered separately PD L1 IHC 22C3 pharmDx GE00621 5 for Dako Omnis Monoclonal Mouse Anti PD L1 Clone 22C3 RTU Dako Omnis with Negative Control Reagent EnVision FLEX High pH Dako Omnis GV80011 2 OR EnVision FLEX Mini Kit High pH Dako Omnis GV82311 2 EnVision FLEX Target Retrieval Solution Low pH 50x GV80511 2 EnVision FLEX Mouse Linker Dako Omnis GV82111 2 EnVision FLEX DAB Enhancer Dako Omnis GC80611 2 PD L1 Control Slides T139130 2 optional The assay requires Low pH Target Retrieval Mouse Linker and a DAB Enhancer but the PD L1 Control Slides are optional For countries outside of the United States see the local KEYTRUDA product label for approved indications and expression cutoff values to guide therapy PD L1 IHC 22C3 pharmDx is subject to an exclusive trademark license to Dako Denmark A S KEYTRUDA is a registered trademark of Merck Sharp Dohme Corp a subsidiary of Merck Co Inc
    https://www.bioz.com/result/egfr/product/Agilent technologies
    Average 97 stars, based on 1 article reviews
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    egfr - by Bioz Stars, 2021-05
    97/100 stars

    Images

    1) Product Images from "Safety and efficacy of afatinib as add-on to standard therapy of gemcitabine/cisplatin in chemotherapy-naive patients with advanced biliary tract cancer: an open-label, phase I trial with an extensive biomarker program"

    Article Title: Safety and efficacy of afatinib as add-on to standard therapy of gemcitabine/cisplatin in chemotherapy-naive patients with advanced biliary tract cancer: an open-label, phase I trial with an extensive biomarker program

    Journal: BMC Cancer

    doi: 10.1186/s12885-018-5223-7

    Immunohistochemical staining, a ) negative control, positive staining of b ) EGFR (3+), c ) HER2neu (1+), d ) HER3 (2+) and e ) HER4 (only sporadic)
    Figure Legend Snippet: Immunohistochemical staining, a ) negative control, positive staining of b ) EGFR (3+), c ) HER2neu (1+), d ) HER3 (2+) and e ) HER4 (only sporadic)

    Techniques Used: Immunohistochemistry, Staining, Negative Control

    2) Product Images from "Identification of EGFR expression status association with metastatic lymph node density (ND) by expression microarray analysis of advanced gastric cancer"

    Article Title: Identification of EGFR expression status association with metastatic lymph node density (ND) by expression microarray analysis of advanced gastric cancer

    Journal: Cancer Medicine

    doi: 10.1002/cam4.311

    High EGFR expression is a strongly associated with high ND. (A) Microscopic analysis of cell membrane EGFR immunohistochemical staining. EGFR expression was graded using a 3-point scale, where 1+ = light staining of more than 10% of the specimens, 2+ = moderate staining of more than 10% and less than or equal to 30% of the specimens, and 3+ = strong staining of more than 30% of the specimens. (B) Statistical analysis of EGFR expression using Student's t -test. EGFR 2+/3+ expression group included more patients with high ND than the EGFR 1+ group. The most suitable ND cutoff level was deemed ND of 35%. (c) Kaplan–Meier curves indicate that EGFR 2+/3+ expression was significantly associated with poor outcome in patients with 13th JGCA stage II/III disease ( P = 0.039). (D) ND ≥35 was significantly associated with poor outcome ( P = 0.0012).
    Figure Legend Snippet: High EGFR expression is a strongly associated with high ND. (A) Microscopic analysis of cell membrane EGFR immunohistochemical staining. EGFR expression was graded using a 3-point scale, where 1+ = light staining of more than 10% of the specimens, 2+ = moderate staining of more than 10% and less than or equal to 30% of the specimens, and 3+ = strong staining of more than 30% of the specimens. (B) Statistical analysis of EGFR expression using Student's t -test. EGFR 2+/3+ expression group included more patients with high ND than the EGFR 1+ group. The most suitable ND cutoff level was deemed ND of 35%. (c) Kaplan–Meier curves indicate that EGFR 2+/3+ expression was significantly associated with poor outcome in patients with 13th JGCA stage II/III disease ( P = 0.039). (D) ND ≥35 was significantly associated with poor outcome ( P = 0.0012).

    Techniques Used: Expressing, Immunohistochemistry, Staining

    3) Product Images from "Establishment and characterization of three stable Basal/HER2-positive breast cancer cell lines derived from Chinese breast carcinoma with identical missense mutations in the DNA-binding domain of TP53"

    Article Title: Establishment and characterization of three stable Basal/HER2-positive breast cancer cell lines derived from Chinese breast carcinoma with identical missense mutations in the DNA-binding domain of TP53

    Journal: Cancer Cell International

    doi: 10.1186/s12935-018-0617-9

    Immunophenotype of ZJU-0327, ZJU-0725 and ZJU-1127 cells in sections from original and xenografted tumors. On the sections of ZJU-0725 and ZJU-1127, strongly positive for ERα, Her2, EGFR, P120 and E-cadherin in original tumors and Her2, EGFR, P120 and E-cadherin in the ZJU-0725 xenografted tumors and Her2, P120 and E-cadherin in the ZJU-0725 xenografted tumors; positive for ki67 in original tumors and ERα and Ki67 in the ZJU-0725 xenografted tumors and ERα, Ki67 and EGFR in the ZJU-1127 xenografted tumors. But negative for PR and ck5/6 in both original and xenografted tumors. On the sections of ZJU-0327, strongly staining for ERα, PR, Her2, P120 and E-cadherin in the original tumors and Her2, P120 and E-cadherin; positive for Ki67 in the original tumors and ERα, ki67 and EGFR. But negative for ck5/6 and EGFR in the original tumors and PR and ck5/6 in the xenografted
    Figure Legend Snippet: Immunophenotype of ZJU-0327, ZJU-0725 and ZJU-1127 cells in sections from original and xenografted tumors. On the sections of ZJU-0725 and ZJU-1127, strongly positive for ERα, Her2, EGFR, P120 and E-cadherin in original tumors and Her2, EGFR, P120 and E-cadherin in the ZJU-0725 xenografted tumors and Her2, P120 and E-cadherin in the ZJU-0725 xenografted tumors; positive for ki67 in original tumors and ERα and Ki67 in the ZJU-0725 xenografted tumors and ERα, Ki67 and EGFR in the ZJU-1127 xenografted tumors. But negative for PR and ck5/6 in both original and xenografted tumors. On the sections of ZJU-0327, strongly staining for ERα, PR, Her2, P120 and E-cadherin in the original tumors and Her2, P120 and E-cadherin; positive for Ki67 in the original tumors and ERα, ki67 and EGFR. But negative for ck5/6 and EGFR in the original tumors and PR and ck5/6 in the xenografted

    Techniques Used: Staining

    4) Product Images from "Immunohistochemical Classification of Primary and Secondary Glioblastomas"

    Article Title: Immunohistochemical Classification of Primary and Secondary Glioblastomas

    Journal: Korean Journal of Pathology

    doi: 10.4132/KoreanJPathol.2013.47.6.541

    Characteristic immunohistochemical expression of epidermal growth factor receptor (EGFR), p53, and isocitrate dehydrogenase 1 (IDH-1) in primary glioblastoma (A-C) and secondary glioblastoma (D-F). EGFR (A, D), p53 (B, E), and IDH-1 (C, F).
    Figure Legend Snippet: Characteristic immunohistochemical expression of epidermal growth factor receptor (EGFR), p53, and isocitrate dehydrogenase 1 (IDH-1) in primary glioblastoma (A-C) and secondary glioblastoma (D-F). EGFR (A, D), p53 (B, E), and IDH-1 (C, F).

    Techniques Used: Immunohistochemistry, Expressing

    5) Product Images from "Distinct gene expression profiles in ovarian cancer linked to Lynch syndrome"

    Article Title: Distinct gene expression profiles in ovarian cancer linked to Lynch syndrome

    Journal: Familial Cancer

    doi: 10.1007/s10689-014-9728-1

    Immunohistochemical stainings for p-mTOR, EGFR and PTEN in ×40 magnification with Lynch syndrome-associated tumors presented on the top row and sporadic tumors on the bottom row. The left and middle columns show positive ( top row ) and negative ( bottom row ) p-mTOR and EGFR expression in tumor cells respectively. The right column shows negative PTEN expression in tumor cells but retained staining in surrounding tissue ( top row ) and positive PTEN expression in tumor cells and surrounding tissue ( bottom row )
    Figure Legend Snippet: Immunohistochemical stainings for p-mTOR, EGFR and PTEN in ×40 magnification with Lynch syndrome-associated tumors presented on the top row and sporadic tumors on the bottom row. The left and middle columns show positive ( top row ) and negative ( bottom row ) p-mTOR and EGFR expression in tumor cells respectively. The right column shows negative PTEN expression in tumor cells but retained staining in surrounding tissue ( top row ) and positive PTEN expression in tumor cells and surrounding tissue ( bottom row )

    Techniques Used: Immunohistochemistry, Expressing, Staining

    6) Product Images from "Prognostic value of expression of molecular markers in adenoid cystic cancer of the salivary glands compared with lymph node metastasis: a retrospective study"

    Article Title: Prognostic value of expression of molecular markers in adenoid cystic cancer of the salivary glands compared with lymph node metastasis: a retrospective study

    Journal: World Journal of Surgical Oncology

    doi: 10.1186/1477-7819-10-266

    Immunohistochemical staining of adenoid cystic carcinomas of the salivary gland. C-kit (CD 117) expression of weakly (1+, A ), moderately (2+, B ), and strongly positive (3+, C ) cases; EGFR (epidermal growth factor receptor) expression of weakly (D) and moderately positive (E) cases; VEGF (vascular endothelial growth factor) expression of weakly (F) , moderately (G) , and strongly positive (H) cases. Expressions of c-kit (membranous/cytoplasmic), EGFR (membranous), and VEGF (cytoplasmic) were scored as follows: 0, reactivity in
    Figure Legend Snippet: Immunohistochemical staining of adenoid cystic carcinomas of the salivary gland. C-kit (CD 117) expression of weakly (1+, A ), moderately (2+, B ), and strongly positive (3+, C ) cases; EGFR (epidermal growth factor receptor) expression of weakly (D) and moderately positive (E) cases; VEGF (vascular endothelial growth factor) expression of weakly (F) , moderately (G) , and strongly positive (H) cases. Expressions of c-kit (membranous/cytoplasmic), EGFR (membranous), and VEGF (cytoplasmic) were scored as follows: 0, reactivity in

    Techniques Used: Immunohistochemistry, Staining, Expressing

    7) Product Images from "Dual targeting of glioblastoma with chimeric antigen receptor-engineered natural killer cells overcomes heterogeneity of target antigen expression and enhances antitumor activity and survival"

    Article Title: Dual targeting of glioblastoma with chimeric antigen receptor-engineered natural killer cells overcomes heterogeneity of target antigen expression and enhances antitumor activity and survival

    Journal: Oncoimmunology

    doi: 10.1080/2162402X.2015.1119354

    Antitumor activity of CAR NK cells against orthotopic LNT-229/EGFR and LNT-229/EGFRvIII GBM xenografts. (A) LNT-229/EGFR cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, or dual-specific NK-92/225 cells once per week for 12 weeks (n = 6). Control mice received injection medium. Tumor growth was monitored by MRI. Tumor development in representative animals from each group at day 53 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). (C) LNT-229/EGFRvIII cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated as described above with NK-92 (n = 6), NK-92/R1 (n = 6), NK-92/MR1-1 (n = 5), or NK-92/225 (n = 6) cells once per week for 8 weeks. Control mice received injection medium (n = 6). Tumor development in representative animals from each group at day 53 is shown. (D) Symptom-free survival of the mice from the experiment described in (C). *** p ≤ 0.001; ** p ≤ 0.01; ns, p > 0.05.
    Figure Legend Snippet: Antitumor activity of CAR NK cells against orthotopic LNT-229/EGFR and LNT-229/EGFRvIII GBM xenografts. (A) LNT-229/EGFR cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, or dual-specific NK-92/225 cells once per week for 12 weeks (n = 6). Control mice received injection medium. Tumor growth was monitored by MRI. Tumor development in representative animals from each group at day 53 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). (C) LNT-229/EGFRvIII cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated as described above with NK-92 (n = 6), NK-92/R1 (n = 6), NK-92/MR1-1 (n = 5), or NK-92/225 (n = 6) cells once per week for 8 weeks. Control mice received injection medium (n = 6). Tumor development in representative animals from each group at day 53 is shown. (D) Symptom-free survival of the mice from the experiment described in (C). *** p ≤ 0.001; ** p ≤ 0.01; ns, p > 0.05.

    Techniques Used: Activity Assay, Injection, Mouse Assay, Magnetic Resonance Imaging

    Generation of CAR NK cells. (A) Lentiviral transfer plasmids pS-R1.28.z-IEW, pS-MR1-1.28.z-IEW and pS-225.28.z-IEW encoding under control of the Spleen Focus Forming Virus promoter (SFFV) CARs consisting of an immunoglobulin heavy chain signal peptide (SP), scFv fragments derived from EGFR-specific antibody R1, EGFRvIII-specific MR1-1, or 225 recognizing EGFR and EGFRvIII, followed by a Myc-tag (M), CD8α hinge region (CD8α), transmembrane and intracellular domains of CD28, and the intracellular domain of CD3ζ. Enhanced green fluorescent protein (EGFP) cDNA separated from the CAR sequence by an internal ribosome entry site (IRES) served as a marker. (B) CAR surface expression on NK-92/R1, NK-92/MR1-1 and NK-92/225 single cell clones was determined by flow cytometry with Myc-tag-specific antibody (open areas). Isotype antibody (filled areas) and parental NK-92 cells served as controls. (C) Binding of recombinant EGFR-Fc protein to the surface of CAR NK cells was measured by flow cytometry (open areas). CAR NK cells only treated with secondary antibody (filled areas) and parental NK-92 cells served as controls. MFI: mean fluorescence intensity (geometric mean).
    Figure Legend Snippet: Generation of CAR NK cells. (A) Lentiviral transfer plasmids pS-R1.28.z-IEW, pS-MR1-1.28.z-IEW and pS-225.28.z-IEW encoding under control of the Spleen Focus Forming Virus promoter (SFFV) CARs consisting of an immunoglobulin heavy chain signal peptide (SP), scFv fragments derived from EGFR-specific antibody R1, EGFRvIII-specific MR1-1, or 225 recognizing EGFR and EGFRvIII, followed by a Myc-tag (M), CD8α hinge region (CD8α), transmembrane and intracellular domains of CD28, and the intracellular domain of CD3ζ. Enhanced green fluorescent protein (EGFP) cDNA separated from the CAR sequence by an internal ribosome entry site (IRES) served as a marker. (B) CAR surface expression on NK-92/R1, NK-92/MR1-1 and NK-92/225 single cell clones was determined by flow cytometry with Myc-tag-specific antibody (open areas). Isotype antibody (filled areas) and parental NK-92 cells served as controls. (C) Binding of recombinant EGFR-Fc protein to the surface of CAR NK cells was measured by flow cytometry (open areas). CAR NK cells only treated with secondary antibody (filled areas) and parental NK-92 cells served as controls. MFI: mean fluorescence intensity (geometric mean).

    Techniques Used: Derivative Assay, Sequencing, Marker, Expressing, Clone Assay, Flow Cytometry, Cytometry, Binding Assay, Recombinant, Fluorescence

    Cytotoxicity of CAR NK cells against EGFR- and EGFRvIII-expressing LNT-229 cells. (A) EGFR (170 kDa) and EGFRvIII (140 kDa) were detected in cell lysates of LNT-229 GBM cells ectopically overexpressing full-length EGFR (LNT-229/EGFR) or mutant EGFRvIII (LNT-229/EGFRvIII) by immunoblotting with an EGFR-specific antibody binding to both receptors. Parental LNT-229 served as control. (B) Cytotoxicity of CAR NK cells against LNT-229/EGFR, LNT-229/EGFRvIII and parental LNT-229 cells was investigated after co-incubation of effector and target cells for 2 h at different E/T ratios. Parental NK-92 were included as control. Mean values ± SEM are shown; n = 3. (C) Conjugate formation between CAR NK cells and LNT-229/EGFR and LNT-229/EGFRvIII cells was investigated by confocal microscopy. Tumor (T) and EGFP-positive CAR NK (N; green) cells were co-incubated for 1 h, fixed, permeabilized and stained for perforin (red) to identify cytotoxic granules. Cell nuclei were labeled with DAPI (blue). Parental NK-92 and NK-92/225.TM cells expressing a CAR without intracellular signaling domains were included as controls. Scale bar: 10 µm.
    Figure Legend Snippet: Cytotoxicity of CAR NK cells against EGFR- and EGFRvIII-expressing LNT-229 cells. (A) EGFR (170 kDa) and EGFRvIII (140 kDa) were detected in cell lysates of LNT-229 GBM cells ectopically overexpressing full-length EGFR (LNT-229/EGFR) or mutant EGFRvIII (LNT-229/EGFRvIII) by immunoblotting with an EGFR-specific antibody binding to both receptors. Parental LNT-229 served as control. (B) Cytotoxicity of CAR NK cells against LNT-229/EGFR, LNT-229/EGFRvIII and parental LNT-229 cells was investigated after co-incubation of effector and target cells for 2 h at different E/T ratios. Parental NK-92 were included as control. Mean values ± SEM are shown; n = 3. (C) Conjugate formation between CAR NK cells and LNT-229/EGFR and LNT-229/EGFRvIII cells was investigated by confocal microscopy. Tumor (T) and EGFP-positive CAR NK (N; green) cells were co-incubated for 1 h, fixed, permeabilized and stained for perforin (red) to identify cytotoxic granules. Cell nuclei were labeled with DAPI (blue). Parental NK-92 and NK-92/225.TM cells expressing a CAR without intracellular signaling domains were included as controls. Scale bar: 10 µm.

    Techniques Used: Expressing, Mutagenesis, Binding Assay, Incubation, Confocal Microscopy, Staining, Labeling

    Antitumor activity of CAR NK cells against mixed LNT-229/EGFR and LNT-229/EGFRvIII GBM xenografts. (A) LNT-229/EGFR and LNT-229/EGFRvIII cells were mixed at a 1:1 ratio before stereotactic injection of the cells into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, dual-specific NK-92/225, or a 1:1 mixture of NK-92/R1 and NK-92/MR1-1 cells once per week for 8 weeks (n = 6). Control mice received injection medium. Tumor development in representative animals from each group at day 54 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). *** p ≤ 0.001; * p ≤ 0.05; ns, p > 0.05. (C) Sections of tumors from individual animals of each treatment group sacrificed at the indicated time point were stained with EGFR- or EGFRvIII-specific antibodies. NK cells present in tumor tissues were detected with CD45-specific antibody. Scale bar: 300 µm.
    Figure Legend Snippet: Antitumor activity of CAR NK cells against mixed LNT-229/EGFR and LNT-229/EGFRvIII GBM xenografts. (A) LNT-229/EGFR and LNT-229/EGFRvIII cells were mixed at a 1:1 ratio before stereotactic injection of the cells into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, dual-specific NK-92/225, or a 1:1 mixture of NK-92/R1 and NK-92/MR1-1 cells once per week for 8 weeks (n = 6). Control mice received injection medium. Tumor development in representative animals from each group at day 54 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). *** p ≤ 0.001; * p ≤ 0.05; ns, p > 0.05. (C) Sections of tumors from individual animals of each treatment group sacrificed at the indicated time point were stained with EGFR- or EGFRvIII-specific antibodies. NK cells present in tumor tissues were detected with CD45-specific antibody. Scale bar: 300 µm.

    Techniques Used: Activity Assay, Injection, Mouse Assay, Staining

    8) Product Images from "Stratification of Prognosis of Triple-Negative Breast Cancer Patients Using Combinatorial Biomarkers"

    Article Title: Stratification of Prognosis of Triple-Negative Breast Cancer Patients Using Combinatorial Biomarkers

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0149661

    Kaplan Meier curves of disease free survival of risk groups for TNBC patients. The patients were stratified into different risk groups. Two low basal (EGFR≤15% and CK5/6≤50%) risk groups: group 1 (n = 20) with low expressions/values of EGFR (≤15%), CK5/6 (≤50%), Ki-67 (≤50%), pT (≤ 3), and pN (≤ 1), and group 2 (n = 30) with low expressions of EGFR and CK5/6, and any high expressions/values of Ki-67, pT, and pN. Two high basal risk groups: group 3 (n = 106) with single high basal expression (EGFR > 15% or CK5/6 > 50%), and group 4 (n = 36) with double high basal expressions (EGFR > 15% and CK5/6 > 50%).
    Figure Legend Snippet: Kaplan Meier curves of disease free survival of risk groups for TNBC patients. The patients were stratified into different risk groups. Two low basal (EGFR≤15% and CK5/6≤50%) risk groups: group 1 (n = 20) with low expressions/values of EGFR (≤15%), CK5/6 (≤50%), Ki-67 (≤50%), pT (≤ 3), and pN (≤ 1), and group 2 (n = 30) with low expressions of EGFR and CK5/6, and any high expressions/values of Ki-67, pT, and pN. Two high basal risk groups: group 3 (n = 106) with single high basal expression (EGFR > 15% or CK5/6 > 50%), and group 4 (n = 36) with double high basal expressions (EGFR > 15% and CK5/6 > 50%).

    Techniques Used: Expressing

    Immunoreactivity of 69-yr-old patient with T1bN0M0 TNBC. (A) Hematoxylin and Eosin (H E), (B) EGFR ( > 70%), (C) CK5/6 ( > 70%), (D) ER, (E) PR, (F) HER2, (G) Ki-67 ( > 60%), and (H) P53 ( > 90%). The patient was undergone BCT, had 3.8 event free survival and 22.8 months overall survival.
    Figure Legend Snippet: Immunoreactivity of 69-yr-old patient with T1bN0M0 TNBC. (A) Hematoxylin and Eosin (H E), (B) EGFR ( > 70%), (C) CK5/6 ( > 70%), (D) ER, (E) PR, (F) HER2, (G) Ki-67 ( > 60%), and (H) P53 ( > 90%). The patient was undergone BCT, had 3.8 event free survival and 22.8 months overall survival.

    Techniques Used:

    Kaplan Meier curves of disease free survival for basal biomarkers. (a) EGFR at cut-off level 15% with log-rank p = 0.0016, (b) time-dependent ROC analysis of EGFR with AUC = 0.723, where cutoff point 15% (red circled), (c) CK5/6 at cut-off level 50% with log-rank p = 0.0066, and (d) the significance of survival difference at different cutoff values for EGFR and CK5/6. The most significant cutoff values were red-circled with 15% for EGFR and 50% for CK5/6.
    Figure Legend Snippet: Kaplan Meier curves of disease free survival for basal biomarkers. (a) EGFR at cut-off level 15% with log-rank p = 0.0016, (b) time-dependent ROC analysis of EGFR with AUC = 0.723, where cutoff point 15% (red circled), (c) CK5/6 at cut-off level 50% with log-rank p = 0.0066, and (d) the significance of survival difference at different cutoff values for EGFR and CK5/6. The most significant cutoff values were red-circled with 15% for EGFR and 50% for CK5/6.

    Techniques Used:

    9) Product Images from "Multiparameter Analysis, including EMT Markers, on Negatively Enriched Blood Samples from Patients with Squamous Cell Carcinoma of the Head and Neck"

    Article Title: Multiparameter Analysis, including EMT Markers, on Negatively Enriched Blood Samples from Patients with Squamous Cell Carcinoma of the Head and Neck

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0042048

    Representative, confocal images of a cytospin of an enriched peripheral blood sample from a SCCHN patient. This is the same patient sample used to create the cytospin presented in Figure 4 . However, for this specific, four color staining, the anti-EGFR antibody was replaced with anti-CD44. The cells highlighted with a red arrow are positive for all four markers, while cells highlighted with yellow arrows are either negative, or very weak for CK, vimentin, and CD44.
    Figure Legend Snippet: Representative, confocal images of a cytospin of an enriched peripheral blood sample from a SCCHN patient. This is the same patient sample used to create the cytospin presented in Figure 4 . However, for this specific, four color staining, the anti-EGFR antibody was replaced with anti-CD44. The cells highlighted with a red arrow are positive for all four markers, while cells highlighted with yellow arrows are either negative, or very weak for CK, vimentin, and CD44.

    Techniques Used: Staining

    Representative, confocal images of a cytospin of an enriched peripheral blood sample from a 27 year old, HPV positive, SCCHN patient. Sample taken in the OR, 214 cytokeratin positive cells per ml of blood were detected, and she had a reoccurrence 2 months after surgery. For this specific four color staining, the cell structure/protein targeted is listed in each image. The cells highlighted with a red arrow are positive for all four markers, yellow are negative for CK, weak or negative for vimentin and EGFR, and white is negative for all but nuclei. Green arrows highlight cells that have a granulocytic like nuclei.
    Figure Legend Snippet: Representative, confocal images of a cytospin of an enriched peripheral blood sample from a 27 year old, HPV positive, SCCHN patient. Sample taken in the OR, 214 cytokeratin positive cells per ml of blood were detected, and she had a reoccurrence 2 months after surgery. For this specific four color staining, the cell structure/protein targeted is listed in each image. The cells highlighted with a red arrow are positive for all four markers, yellow are negative for CK, weak or negative for vimentin and EGFR, and white is negative for all but nuclei. Green arrows highlight cells that have a granulocytic like nuclei.

    Techniques Used: Staining

    Representative set of confocal images of a cytospin of stained normal blood samples and SSC4 cell lines. The cell structure/protein targeted is listed in each image, the type of cells listed on right. Note, the second row of images presents cells stained for EGFR, while the third row shows cells stained for CD44.
    Figure Legend Snippet: Representative set of confocal images of a cytospin of stained normal blood samples and SSC4 cell lines. The cell structure/protein targeted is listed in each image, the type of cells listed on right. Note, the second row of images presents cells stained for EGFR, while the third row shows cells stained for CD44.

    Techniques Used: Staining

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    Fluorescence In Situ Hybridization:

    Article Title: PD‐L1 expression in ROS1‐rearranged non‐small cell lung cancer: A study using simultaneous genotypic screening of EGFR, ALK, and ROS1
    Article Snippet: .. Additionally, the sample needed to have > 100 tumor cells for the PD‐L1 test and > 50 tumors cells for ALK and ROS1 FISH tests., In a tissue‐sparing manner, we also investigated an IHC panel including thyroid transcription factor 1 (TTF‐1), p63, p40, and cytokeratin 7; mucin staining using Periodic acid‐Schiff stain and Alcian blue for histological confirmation; ALK IHC to confirm the FISH results; and PD‐L1 IHC (using the PD‐L1 IHC 22C3 pharmDx, Agilent/Dako) for therapeutic purposes., PD‐L1 IHC was performed on 4 μm thick formalin‐fixed paraffin‐embedded (FFPE) tissue sections using the PD‐L1 clone 22C3 pharmDx kit and Dako Automated Link 48 platform (Agilent/Dako). .. Immunohistochemical staining for TTF‐1 (clone SP141, Ventana Medical Systems, Tucson, AZ, USA), p63 (clone 4A4), p40 (polyclonal anti p40; Diagnostic BioSystems, Pleasanton, CA, USA), cytokeratin 7 (clone SP52, Ventana), and ALK (clone 5A4, dilution 1:50, Leica Biosystems, Newcastle, UK) was performed using the Benchmark Ultra Autostainer (Roche Tissue Diagnostics, Tucson, AZ, USA) with an OptiView Universal DAB detection kit for ALK IHC and UltraView universal DAB detection kit for the other proteins (Ventana Medical Systems).

    Immunohistochemistry:

    Article Title: PD‐L1 expression in ROS1‐rearranged non‐small cell lung cancer: A study using simultaneous genotypic screening of EGFR, ALK, and ROS1
    Article Snippet: .. Additionally, the sample needed to have > 100 tumor cells for the PD‐L1 test and > 50 tumors cells for ALK and ROS1 FISH tests., In a tissue‐sparing manner, we also investigated an IHC panel including thyroid transcription factor 1 (TTF‐1), p63, p40, and cytokeratin 7; mucin staining using Periodic acid‐Schiff stain and Alcian blue for histological confirmation; ALK IHC to confirm the FISH results; and PD‐L1 IHC (using the PD‐L1 IHC 22C3 pharmDx, Agilent/Dako) for therapeutic purposes., PD‐L1 IHC was performed on 4 μm thick formalin‐fixed paraffin‐embedded (FFPE) tissue sections using the PD‐L1 clone 22C3 pharmDx kit and Dako Automated Link 48 platform (Agilent/Dako). .. Immunohistochemical staining for TTF‐1 (clone SP141, Ventana Medical Systems, Tucson, AZ, USA), p63 (clone 4A4), p40 (polyclonal anti p40; Diagnostic BioSystems, Pleasanton, CA, USA), cytokeratin 7 (clone SP52, Ventana), and ALK (clone 5A4, dilution 1:50, Leica Biosystems, Newcastle, UK) was performed using the Benchmark Ultra Autostainer (Roche Tissue Diagnostics, Tucson, AZ, USA) with an OptiView Universal DAB detection kit for ALK IHC and UltraView universal DAB detection kit for the other proteins (Ventana Medical Systems).

    Article Title: Immunohistochemical assays incorporating SP142 and 22C3 monoclonal antibodies for detection of PD-L1 expression in NSCLC patients with known status of EGFR and ALK genes
    Article Snippet: .. We compared the effectiveness of PD-L1 expression examination of two IHC assays with 22C3 (Dako) and SP142 antibodies (Ventana). .. IHC tests were performed in resected tissue samples and in cellblocks from bronchoscopy biopsies (formalin-fixed paraffin-embedded).

    Article Title: Multiple lung cancers including squamous cell carcinoma with strong PD-L1 expression and adenocarcinoma with EGFR exon 19 deletion: A case report
    Article Snippet: .. SQCC of the right middle lobe showed 100% tumor proportion score (TPS) for PD-L1 (Agilent Dako IHC 22C3 platform) ( B) and no expression of EGFR mutations (Roche cobas® EGFR Mutation Test v2) and anaplastic lymphoma kinase (ALK) rearrangements (Histofine ALK iAEP® Kit). .. Adenocarcinoma of the right lower lobe showed exon 19 deletion and no expression of PD-L1.

    Article Title: Any Place for Immunohistochemistry within the Predictive Biomarkers of Treatment in Lung Cancer Patients?
    Article Snippet: .. The 22C3 test (PD-L1 IHC 22C3 pharmDx, Agilent Technologies, Inc., Santa Clara, CA, USA) is currently the only test used as a companion diagnostic test (CDX) for the administration of pembrolizumab as first line treatement in advanced or metastatic NSCLC [ ]. ..

    Article Title: PD-L1 Expression Correlated with p53 Expression in Pediatric Glioblastoma Multiforme
    Article Snippet: The analysis was performed on a Dako Omnis IHC platform (GI100, Santa Clara, CA, USA). .. Selected reagents were used in the IHC analysis: PD-L1 IHC 22C3 pharmDx staining set (GE006) including monoclonal PD-L1 antibody 22C3 clone (Dako Omnis, Santa Clara, CA, USA), negative control (Dako Omnis, Santa Clara, CA, USA), high pH detective sys- tem EnVision Flex Mini Kit (Dako Omnis, Santa Clara, CA, USA, GV823), wash buffer (20x) (GC807, Dako Omnis, Santa Clara, CA, USA). .. In order to remove the paraffin, a low pH Envision Flex Target Retrieval Solution (Dako Omnis, Santa Clara, CA, USA) was used.

    Article Title: Evaluation of an online training tool for scoring programmed cell death ligand-1 (PD-L1) diagnostic tests for lung cancer
    Article Snippet: In the competence test set, the consensus scores for PD-L1 TC expression for samples stained using the VENTANA PD-L1 (SP263) assay were: 3 cases < 1%; 3 cases ≥1– < 25%; 3 cases ≥25– < 50%; 9 cases ≥50%. .. For samples stained using the Dako PD-L1 IHC PharmDx 22C3 assay, expression was classified as follows: 2 cases < 1%; 6 cases ≥1– < 25%; 4 cases ≥25– < 50%; 6 cases ≥50%. ..

    Staining:

    Article Title: PD‐L1 expression in ROS1‐rearranged non‐small cell lung cancer: A study using simultaneous genotypic screening of EGFR, ALK, and ROS1
    Article Snippet: .. Additionally, the sample needed to have > 100 tumor cells for the PD‐L1 test and > 50 tumors cells for ALK and ROS1 FISH tests., In a tissue‐sparing manner, we also investigated an IHC panel including thyroid transcription factor 1 (TTF‐1), p63, p40, and cytokeratin 7; mucin staining using Periodic acid‐Schiff stain and Alcian blue for histological confirmation; ALK IHC to confirm the FISH results; and PD‐L1 IHC (using the PD‐L1 IHC 22C3 pharmDx, Agilent/Dako) for therapeutic purposes., PD‐L1 IHC was performed on 4 μm thick formalin‐fixed paraffin‐embedded (FFPE) tissue sections using the PD‐L1 clone 22C3 pharmDx kit and Dako Automated Link 48 platform (Agilent/Dako). .. Immunohistochemical staining for TTF‐1 (clone SP141, Ventana Medical Systems, Tucson, AZ, USA), p63 (clone 4A4), p40 (polyclonal anti p40; Diagnostic BioSystems, Pleasanton, CA, USA), cytokeratin 7 (clone SP52, Ventana), and ALK (clone 5A4, dilution 1:50, Leica Biosystems, Newcastle, UK) was performed using the Benchmark Ultra Autostainer (Roche Tissue Diagnostics, Tucson, AZ, USA) with an OptiView Universal DAB detection kit for ALK IHC and UltraView universal DAB detection kit for the other proteins (Ventana Medical Systems).

    Article Title: PD-L1 Expression Correlated with p53 Expression in Pediatric Glioblastoma Multiforme
    Article Snippet: The analysis was performed on a Dako Omnis IHC platform (GI100, Santa Clara, CA, USA). .. Selected reagents were used in the IHC analysis: PD-L1 IHC 22C3 pharmDx staining set (GE006) including monoclonal PD-L1 antibody 22C3 clone (Dako Omnis, Santa Clara, CA, USA), negative control (Dako Omnis, Santa Clara, CA, USA), high pH detective sys- tem EnVision Flex Mini Kit (Dako Omnis, Santa Clara, CA, USA, GV823), wash buffer (20x) (GC807, Dako Omnis, Santa Clara, CA, USA). .. In order to remove the paraffin, a low pH Envision Flex Target Retrieval Solution (Dako Omnis, Santa Clara, CA, USA) was used.

    Article Title: Evaluation of an online training tool for scoring programmed cell death ligand-1 (PD-L1) diagnostic tests for lung cancer
    Article Snippet: In the competence test set, the consensus scores for PD-L1 TC expression for samples stained using the VENTANA PD-L1 (SP263) assay were: 3 cases < 1%; 3 cases ≥1– < 25%; 3 cases ≥25– < 50%; 9 cases ≥50%. .. For samples stained using the Dako PD-L1 IHC PharmDx 22C3 assay, expression was classified as follows: 2 cases < 1%; 6 cases ≥1– < 25%; 4 cases ≥25– < 50%; 6 cases ≥50%. ..

    Formalin-fixed Paraffin-Embedded:

    Article Title: PD‐L1 expression in ROS1‐rearranged non‐small cell lung cancer: A study using simultaneous genotypic screening of EGFR, ALK, and ROS1
    Article Snippet: .. Additionally, the sample needed to have > 100 tumor cells for the PD‐L1 test and > 50 tumors cells for ALK and ROS1 FISH tests., In a tissue‐sparing manner, we also investigated an IHC panel including thyroid transcription factor 1 (TTF‐1), p63, p40, and cytokeratin 7; mucin staining using Periodic acid‐Schiff stain and Alcian blue for histological confirmation; ALK IHC to confirm the FISH results; and PD‐L1 IHC (using the PD‐L1 IHC 22C3 pharmDx, Agilent/Dako) for therapeutic purposes., PD‐L1 IHC was performed on 4 μm thick formalin‐fixed paraffin‐embedded (FFPE) tissue sections using the PD‐L1 clone 22C3 pharmDx kit and Dako Automated Link 48 platform (Agilent/Dako). .. Immunohistochemical staining for TTF‐1 (clone SP141, Ventana Medical Systems, Tucson, AZ, USA), p63 (clone 4A4), p40 (polyclonal anti p40; Diagnostic BioSystems, Pleasanton, CA, USA), cytokeratin 7 (clone SP52, Ventana), and ALK (clone 5A4, dilution 1:50, Leica Biosystems, Newcastle, UK) was performed using the Benchmark Ultra Autostainer (Roche Tissue Diagnostics, Tucson, AZ, USA) with an OptiView Universal DAB detection kit for ALK IHC and UltraView universal DAB detection kit for the other proteins (Ventana Medical Systems).

    Expressing:

    Article Title: Immunohistochemical assays incorporating SP142 and 22C3 monoclonal antibodies for detection of PD-L1 expression in NSCLC patients with known status of EGFR and ALK genes
    Article Snippet: .. We compared the effectiveness of PD-L1 expression examination of two IHC assays with 22C3 (Dako) and SP142 antibodies (Ventana). .. IHC tests were performed in resected tissue samples and in cellblocks from bronchoscopy biopsies (formalin-fixed paraffin-embedded).

    Article Title: Multiple lung cancers including squamous cell carcinoma with strong PD-L1 expression and adenocarcinoma with EGFR exon 19 deletion: A case report
    Article Snippet: .. SQCC of the right middle lobe showed 100% tumor proportion score (TPS) for PD-L1 (Agilent Dako IHC 22C3 platform) ( B) and no expression of EGFR mutations (Roche cobas® EGFR Mutation Test v2) and anaplastic lymphoma kinase (ALK) rearrangements (Histofine ALK iAEP® Kit). .. Adenocarcinoma of the right lower lobe showed exon 19 deletion and no expression of PD-L1.

    Article Title: Evaluation of an online training tool for scoring programmed cell death ligand-1 (PD-L1) diagnostic tests for lung cancer
    Article Snippet: In the competence test set, the consensus scores for PD-L1 TC expression for samples stained using the VENTANA PD-L1 (SP263) assay were: 3 cases < 1%; 3 cases ≥1– < 25%; 3 cases ≥25– < 50%; 9 cases ≥50%. .. For samples stained using the Dako PD-L1 IHC PharmDx 22C3 assay, expression was classified as follows: 2 cases < 1%; 6 cases ≥1– < 25%; 4 cases ≥25– < 50%; 6 cases ≥50%. ..

    Mutagenesis:

    Article Title: Multiple lung cancers including squamous cell carcinoma with strong PD-L1 expression and adenocarcinoma with EGFR exon 19 deletion: A case report
    Article Snippet: .. SQCC of the right middle lobe showed 100% tumor proportion score (TPS) for PD-L1 (Agilent Dako IHC 22C3 platform) ( B) and no expression of EGFR mutations (Roche cobas® EGFR Mutation Test v2) and anaplastic lymphoma kinase (ALK) rearrangements (Histofine ALK iAEP® Kit). .. Adenocarcinoma of the right lower lobe showed exon 19 deletion and no expression of PD-L1.

    Diagnostic Assay:

    Article Title: Any Place for Immunohistochemistry within the Predictive Biomarkers of Treatment in Lung Cancer Patients?
    Article Snippet: .. The 22C3 test (PD-L1 IHC 22C3 pharmDx, Agilent Technologies, Inc., Santa Clara, CA, USA) is currently the only test used as a companion diagnostic test (CDX) for the administration of pembrolizumab as first line treatement in advanced or metastatic NSCLC [ ]. ..

    Article Title: Use of the 22C3 anti–PD-L1 antibody to determine PD-L1 expression in multiple automated immunohistochemistry platforms
    Article Snippet: In patients with metastatic NSCLC and no prior systemic therapy, pembrolizumab was recently approved in the United States for the treatment of patients with PD-L1 expression ≥50% based on the results of the KEYNOTE-024 (NCT02142738) study [ ]. .. Based on these data, pembrolizumab was approved in conjunction with a companion diagnostic test, the PD-L1 IHC 22C3 pharmDx assay (Dako, Carpinteria, CA) for use on the Dako Autostainer Link 48 (ASL48) platform [ , ]. .. However, pathology laboratories without the ASL48 platform are currently unable to provide PD-L1 immunohistochemistry (IHC) staining to identify patients with NSCLC suitable for treatment with pembrolizumab.

    Negative Control:

    Article Title: PD-L1 Expression Correlated with p53 Expression in Pediatric Glioblastoma Multiforme
    Article Snippet: The analysis was performed on a Dako Omnis IHC platform (GI100, Santa Clara, CA, USA). .. Selected reagents were used in the IHC analysis: PD-L1 IHC 22C3 pharmDx staining set (GE006) including monoclonal PD-L1 antibody 22C3 clone (Dako Omnis, Santa Clara, CA, USA), negative control (Dako Omnis, Santa Clara, CA, USA), high pH detective sys- tem EnVision Flex Mini Kit (Dako Omnis, Santa Clara, CA, USA, GV823), wash buffer (20x) (GC807, Dako Omnis, Santa Clara, CA, USA). .. In order to remove the paraffin, a low pH Envision Flex Target Retrieval Solution (Dako Omnis, Santa Clara, CA, USA) was used.

    Transmission Electron Microscopy:

    Article Title: PD-L1 Expression Correlated with p53 Expression in Pediatric Glioblastoma Multiforme
    Article Snippet: The analysis was performed on a Dako Omnis IHC platform (GI100, Santa Clara, CA, USA). .. Selected reagents were used in the IHC analysis: PD-L1 IHC 22C3 pharmDx staining set (GE006) including monoclonal PD-L1 antibody 22C3 clone (Dako Omnis, Santa Clara, CA, USA), negative control (Dako Omnis, Santa Clara, CA, USA), high pH detective sys- tem EnVision Flex Mini Kit (Dako Omnis, Santa Clara, CA, USA, GV823), wash buffer (20x) (GC807, Dako Omnis, Santa Clara, CA, USA). .. In order to remove the paraffin, a low pH Envision Flex Target Retrieval Solution (Dako Omnis, Santa Clara, CA, USA) was used.

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    Agilent technologies egfr
    Antitumor activity of CAR NK cells against orthotopic <t>LNT-229/EGFR</t> and <t>LNT-229/EGFRvIII</t> GBM xenografts. (A) LNT-229/EGFR cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, or dual-specific NK-92/225 cells once per week for 12 weeks (n = 6). Control mice received injection medium. Tumor growth was monitored by MRI. Tumor development in representative animals from each group at day 53 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). (C) LNT-229/EGFRvIII cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated as described above with NK-92 (n = 6), NK-92/R1 (n = 6), NK-92/MR1-1 (n = 5), or NK-92/225 (n = 6) cells once per week for 8 weeks. Control mice received injection medium (n = 6). Tumor development in representative animals from each group at day 53 is shown. (D) Symptom-free survival of the mice from the experiment described in (C). *** p ≤ 0.001; ** p ≤ 0.01; ns, p > 0.05.
    Egfr, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Agilent technologies human epidermal growth factor receptor egfr
    High <t>EGFR</t> expression is a strongly associated with high ND. (A) Microscopic analysis of cell membrane EGFR <t>immunohistochemical</t> staining. EGFR expression was graded using a 3-point scale, where 1+ = light staining of more than 10% of the specimens, 2+ = moderate staining of more than 10% and less than or equal to 30% of the specimens, and 3+ = strong staining of more than 30% of the specimens. (B) Statistical analysis of EGFR expression using Student's t -test. EGFR 2+/3+ expression group included more patients with high ND than the EGFR 1+ group. The most suitable ND cutoff level was deemed ND of 35%. (c) Kaplan–Meier curves indicate that EGFR 2+/3+ expression was significantly associated with poor outcome in patients with 13th JGCA stage II/III disease ( P = 0.039). (D) ND ≥35 was significantly associated with poor outcome ( P = 0.0012).
    Human Epidermal Growth Factor Receptor Egfr, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies egfr expression
    FISH analysis with two different <t>EGFR-specific</t> FISH probes. A , B , C , D : <t>Dako</t> Cytomation FISH probe mix (EGFR: red, CEN7: green), E , F , G , H : ZytoLight SPEC EGFR/CEN7 dual probe (EGFR: green, CEN7: red), (magnification × 630) A , E : balanced disomy, B , F : balanced trisomy, C , G : low amplification, D , H : high amplification.
    Egfr Expression, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies mouse anti human egfr monoclonal antibody
    <t>EGFR</t> (epidermal growth factor receptor) gene status in adrenocortical neoplasms: (a) This image demonstrates strong membrane EGFR expression (3+) in adrenocortical carcinoma, as detected by <t>immunohistochemistry.</t> (b) This image demonstrates an absence of membrane EGFR expression in adrenocortical adenoma, as detected by immunohistochemistry. (c) The cancer cells demonstrate high polysomy on chromosome 7 in adrenocortical carcinoma, as detected by FISH(fluorescence in situ hybridization). (d) The tumor cells display disomy for the EGFR in adrenocortical adenoma, as detected by FISH (Green signals represent the chromosome 7 centromere, and red signals represent the EGFR gene).
    Mouse Anti Human Egfr Monoclonal Antibody, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Antitumor activity of CAR NK cells against orthotopic LNT-229/EGFR and LNT-229/EGFRvIII GBM xenografts. (A) LNT-229/EGFR cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, or dual-specific NK-92/225 cells once per week for 12 weeks (n = 6). Control mice received injection medium. Tumor growth was monitored by MRI. Tumor development in representative animals from each group at day 53 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). (C) LNT-229/EGFRvIII cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated as described above with NK-92 (n = 6), NK-92/R1 (n = 6), NK-92/MR1-1 (n = 5), or NK-92/225 (n = 6) cells once per week for 8 weeks. Control mice received injection medium (n = 6). Tumor development in representative animals from each group at day 53 is shown. (D) Symptom-free survival of the mice from the experiment described in (C). *** p ≤ 0.001; ** p ≤ 0.01; ns, p > 0.05.

    Journal: Oncoimmunology

    Article Title: Dual targeting of glioblastoma with chimeric antigen receptor-engineered natural killer cells overcomes heterogeneity of target antigen expression and enhances antitumor activity and survival

    doi: 10.1080/2162402X.2015.1119354

    Figure Lengend Snippet: Antitumor activity of CAR NK cells against orthotopic LNT-229/EGFR and LNT-229/EGFRvIII GBM xenografts. (A) LNT-229/EGFR cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, or dual-specific NK-92/225 cells once per week for 12 weeks (n = 6). Control mice received injection medium. Tumor growth was monitored by MRI. Tumor development in representative animals from each group at day 53 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). (C) LNT-229/EGFRvIII cells were stereotactically injected into the right striatum of NSG mice. Seven days later, the animals were treated as described above with NK-92 (n = 6), NK-92/R1 (n = 6), NK-92/MR1-1 (n = 5), or NK-92/225 (n = 6) cells once per week for 8 weeks. Control mice received injection medium (n = 6). Tumor development in representative animals from each group at day 53 is shown. (D) Symptom-free survival of the mice from the experiment described in (C). *** p ≤ 0.001; ** p ≤ 0.01; ns, p > 0.05.

    Article Snippet: Slides were stained according to a standardized staining protocol (Bond Polymer Refine IHC protocol, IHC-F; Leica Microsystems) using murine primary antibodies specific for human CD45 (clone 2B11 + PD7/26, 1:100 dilution; Dako), EGFR (clone DAK-H1-WT, 1:50 dilution; Dako) and EGFRvIII (DH8.3, 14 µg/mL final concentration), followed by polymeric HRP-conjugated anti-mouse antibody (DAB Polymer Refine Detection Kit; Leica Microsystems).

    Techniques: Activity Assay, Injection, Mouse Assay, Magnetic Resonance Imaging

    Generation of CAR NK cells. (A) Lentiviral transfer plasmids pS-R1.28.z-IEW, pS-MR1-1.28.z-IEW and pS-225.28.z-IEW encoding under control of the Spleen Focus Forming Virus promoter (SFFV) CARs consisting of an immunoglobulin heavy chain signal peptide (SP), scFv fragments derived from EGFR-specific antibody R1, EGFRvIII-specific MR1-1, or 225 recognizing EGFR and EGFRvIII, followed by a Myc-tag (M), CD8α hinge region (CD8α), transmembrane and intracellular domains of CD28, and the intracellular domain of CD3ζ. Enhanced green fluorescent protein (EGFP) cDNA separated from the CAR sequence by an internal ribosome entry site (IRES) served as a marker. (B) CAR surface expression on NK-92/R1, NK-92/MR1-1 and NK-92/225 single cell clones was determined by flow cytometry with Myc-tag-specific antibody (open areas). Isotype antibody (filled areas) and parental NK-92 cells served as controls. (C) Binding of recombinant EGFR-Fc protein to the surface of CAR NK cells was measured by flow cytometry (open areas). CAR NK cells only treated with secondary antibody (filled areas) and parental NK-92 cells served as controls. MFI: mean fluorescence intensity (geometric mean).

    Journal: Oncoimmunology

    Article Title: Dual targeting of glioblastoma with chimeric antigen receptor-engineered natural killer cells overcomes heterogeneity of target antigen expression and enhances antitumor activity and survival

    doi: 10.1080/2162402X.2015.1119354

    Figure Lengend Snippet: Generation of CAR NK cells. (A) Lentiviral transfer plasmids pS-R1.28.z-IEW, pS-MR1-1.28.z-IEW and pS-225.28.z-IEW encoding under control of the Spleen Focus Forming Virus promoter (SFFV) CARs consisting of an immunoglobulin heavy chain signal peptide (SP), scFv fragments derived from EGFR-specific antibody R1, EGFRvIII-specific MR1-1, or 225 recognizing EGFR and EGFRvIII, followed by a Myc-tag (M), CD8α hinge region (CD8α), transmembrane and intracellular domains of CD28, and the intracellular domain of CD3ζ. Enhanced green fluorescent protein (EGFP) cDNA separated from the CAR sequence by an internal ribosome entry site (IRES) served as a marker. (B) CAR surface expression on NK-92/R1, NK-92/MR1-1 and NK-92/225 single cell clones was determined by flow cytometry with Myc-tag-specific antibody (open areas). Isotype antibody (filled areas) and parental NK-92 cells served as controls. (C) Binding of recombinant EGFR-Fc protein to the surface of CAR NK cells was measured by flow cytometry (open areas). CAR NK cells only treated with secondary antibody (filled areas) and parental NK-92 cells served as controls. MFI: mean fluorescence intensity (geometric mean).

    Article Snippet: Slides were stained according to a standardized staining protocol (Bond Polymer Refine IHC protocol, IHC-F; Leica Microsystems) using murine primary antibodies specific for human CD45 (clone 2B11 + PD7/26, 1:100 dilution; Dako), EGFR (clone DAK-H1-WT, 1:50 dilution; Dako) and EGFRvIII (DH8.3, 14 µg/mL final concentration), followed by polymeric HRP-conjugated anti-mouse antibody (DAB Polymer Refine Detection Kit; Leica Microsystems).

    Techniques: Derivative Assay, Sequencing, Marker, Expressing, Clone Assay, Flow Cytometry, Cytometry, Binding Assay, Recombinant, Fluorescence

    Cytotoxicity of CAR NK cells against EGFR- and EGFRvIII-expressing LNT-229 cells. (A) EGFR (170 kDa) and EGFRvIII (140 kDa) were detected in cell lysates of LNT-229 GBM cells ectopically overexpressing full-length EGFR (LNT-229/EGFR) or mutant EGFRvIII (LNT-229/EGFRvIII) by immunoblotting with an EGFR-specific antibody binding to both receptors. Parental LNT-229 served as control. (B) Cytotoxicity of CAR NK cells against LNT-229/EGFR, LNT-229/EGFRvIII and parental LNT-229 cells was investigated after co-incubation of effector and target cells for 2 h at different E/T ratios. Parental NK-92 were included as control. Mean values ± SEM are shown; n = 3. (C) Conjugate formation between CAR NK cells and LNT-229/EGFR and LNT-229/EGFRvIII cells was investigated by confocal microscopy. Tumor (T) and EGFP-positive CAR NK (N; green) cells were co-incubated for 1 h, fixed, permeabilized and stained for perforin (red) to identify cytotoxic granules. Cell nuclei were labeled with DAPI (blue). Parental NK-92 and NK-92/225.TM cells expressing a CAR without intracellular signaling domains were included as controls. Scale bar: 10 µm.

    Journal: Oncoimmunology

    Article Title: Dual targeting of glioblastoma with chimeric antigen receptor-engineered natural killer cells overcomes heterogeneity of target antigen expression and enhances antitumor activity and survival

    doi: 10.1080/2162402X.2015.1119354

    Figure Lengend Snippet: Cytotoxicity of CAR NK cells against EGFR- and EGFRvIII-expressing LNT-229 cells. (A) EGFR (170 kDa) and EGFRvIII (140 kDa) were detected in cell lysates of LNT-229 GBM cells ectopically overexpressing full-length EGFR (LNT-229/EGFR) or mutant EGFRvIII (LNT-229/EGFRvIII) by immunoblotting with an EGFR-specific antibody binding to both receptors. Parental LNT-229 served as control. (B) Cytotoxicity of CAR NK cells against LNT-229/EGFR, LNT-229/EGFRvIII and parental LNT-229 cells was investigated after co-incubation of effector and target cells for 2 h at different E/T ratios. Parental NK-92 were included as control. Mean values ± SEM are shown; n = 3. (C) Conjugate formation between CAR NK cells and LNT-229/EGFR and LNT-229/EGFRvIII cells was investigated by confocal microscopy. Tumor (T) and EGFP-positive CAR NK (N; green) cells were co-incubated for 1 h, fixed, permeabilized and stained for perforin (red) to identify cytotoxic granules. Cell nuclei were labeled with DAPI (blue). Parental NK-92 and NK-92/225.TM cells expressing a CAR without intracellular signaling domains were included as controls. Scale bar: 10 µm.

    Article Snippet: Slides were stained according to a standardized staining protocol (Bond Polymer Refine IHC protocol, IHC-F; Leica Microsystems) using murine primary antibodies specific for human CD45 (clone 2B11 + PD7/26, 1:100 dilution; Dako), EGFR (clone DAK-H1-WT, 1:50 dilution; Dako) and EGFRvIII (DH8.3, 14 µg/mL final concentration), followed by polymeric HRP-conjugated anti-mouse antibody (DAB Polymer Refine Detection Kit; Leica Microsystems).

    Techniques: Expressing, Mutagenesis, Binding Assay, Incubation, Confocal Microscopy, Staining, Labeling

    Antitumor activity of CAR NK cells against mixed LNT-229/EGFR and LNT-229/EGFRvIII GBM xenografts. (A) LNT-229/EGFR and LNT-229/EGFRvIII cells were mixed at a 1:1 ratio before stereotactic injection of the cells into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, dual-specific NK-92/225, or a 1:1 mixture of NK-92/R1 and NK-92/MR1-1 cells once per week for 8 weeks (n = 6). Control mice received injection medium. Tumor development in representative animals from each group at day 54 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). *** p ≤ 0.001; * p ≤ 0.05; ns, p > 0.05. (C) Sections of tumors from individual animals of each treatment group sacrificed at the indicated time point were stained with EGFR- or EGFRvIII-specific antibodies. NK cells present in tumor tissues were detected with CD45-specific antibody. Scale bar: 300 µm.

    Journal: Oncoimmunology

    Article Title: Dual targeting of glioblastoma with chimeric antigen receptor-engineered natural killer cells overcomes heterogeneity of target antigen expression and enhances antitumor activity and survival

    doi: 10.1080/2162402X.2015.1119354

    Figure Lengend Snippet: Antitumor activity of CAR NK cells against mixed LNT-229/EGFR and LNT-229/EGFRvIII GBM xenografts. (A) LNT-229/EGFR and LNT-229/EGFRvIII cells were mixed at a 1:1 ratio before stereotactic injection of the cells into the right striatum of NSG mice. Seven days later, the animals were treated by intratumoral injection of parental NK-92, EGFR-specific NK-92/R1, EGFRvIII-specific NK-92/MR1-1, dual-specific NK-92/225, or a 1:1 mixture of NK-92/R1 and NK-92/MR1-1 cells once per week for 8 weeks (n = 6). Control mice received injection medium. Tumor development in representative animals from each group at day 54 is shown. (B) Symptom-free survival of the mice from the experiment described in (A). *** p ≤ 0.001; * p ≤ 0.05; ns, p > 0.05. (C) Sections of tumors from individual animals of each treatment group sacrificed at the indicated time point were stained with EGFR- or EGFRvIII-specific antibodies. NK cells present in tumor tissues were detected with CD45-specific antibody. Scale bar: 300 µm.

    Article Snippet: Slides were stained according to a standardized staining protocol (Bond Polymer Refine IHC protocol, IHC-F; Leica Microsystems) using murine primary antibodies specific for human CD45 (clone 2B11 + PD7/26, 1:100 dilution; Dako), EGFR (clone DAK-H1-WT, 1:50 dilution; Dako) and EGFRvIII (DH8.3, 14 µg/mL final concentration), followed by polymeric HRP-conjugated anti-mouse antibody (DAB Polymer Refine Detection Kit; Leica Microsystems).

    Techniques: Activity Assay, Injection, Mouse Assay, Staining

    High EGFR expression is a strongly associated with high ND. (A) Microscopic analysis of cell membrane EGFR immunohistochemical staining. EGFR expression was graded using a 3-point scale, where 1+ = light staining of more than 10% of the specimens, 2+ = moderate staining of more than 10% and less than or equal to 30% of the specimens, and 3+ = strong staining of more than 30% of the specimens. (B) Statistical analysis of EGFR expression using Student's t -test. EGFR 2+/3+ expression group included more patients with high ND than the EGFR 1+ group. The most suitable ND cutoff level was deemed ND of 35%. (c) Kaplan–Meier curves indicate that EGFR 2+/3+ expression was significantly associated with poor outcome in patients with 13th JGCA stage II/III disease ( P = 0.039). (D) ND ≥35 was significantly associated with poor outcome ( P = 0.0012).

    Journal: Cancer Medicine

    Article Title: Identification of EGFR expression status association with metastatic lymph node density (ND) by expression microarray analysis of advanced gastric cancer

    doi: 10.1002/cam4.311

    Figure Lengend Snippet: High EGFR expression is a strongly associated with high ND. (A) Microscopic analysis of cell membrane EGFR immunohistochemical staining. EGFR expression was graded using a 3-point scale, where 1+ = light staining of more than 10% of the specimens, 2+ = moderate staining of more than 10% and less than or equal to 30% of the specimens, and 3+ = strong staining of more than 30% of the specimens. (B) Statistical analysis of EGFR expression using Student's t -test. EGFR 2+/3+ expression group included more patients with high ND than the EGFR 1+ group. The most suitable ND cutoff level was deemed ND of 35%. (c) Kaplan–Meier curves indicate that EGFR 2+/3+ expression was significantly associated with poor outcome in patients with 13th JGCA stage II/III disease ( P = 0.039). (D) ND ≥35 was significantly associated with poor outcome ( P = 0.0012).

    Article Snippet: Immunohistochemical staining of the EGFR The primary antibodies used for immunohistochemical (IHC) assays were the mouse monoclonal antibodies against the human epidermal growth factor receptor (EGFR) that are included in the EGFR pharmDx kit (Dako-Japan, Tokyo, Japan).

    Techniques: Expressing, Immunohistochemistry, Staining

    FISH analysis with two different EGFR-specific FISH probes. A , B , C , D : Dako Cytomation FISH probe mix (EGFR: red, CEN7: green), E , F , G , H : ZytoLight SPEC EGFR/CEN7 dual probe (EGFR: green, CEN7: red), (magnification × 630) A , E : balanced disomy, B , F : balanced trisomy, C , G : low amplification, D , H : high amplification.

    Journal: Diagnostic Pathology

    Article Title: Correlation of EGFR expression, gene copy number and clinicopathological status in NSCLC

    doi: 10.1186/s13000-014-0165-0

    Figure Lengend Snippet: FISH analysis with two different EGFR-specific FISH probes. A , B , C , D : Dako Cytomation FISH probe mix (EGFR: red, CEN7: green), E , F , G , H : ZytoLight SPEC EGFR/CEN7 dual probe (EGFR: green, CEN7: red), (magnification × 630) A , E : balanced disomy, B , F : balanced trisomy, C , G : low amplification, D , H : high amplification.

    Article Snippet: Scoring with method (B) showed a similar EGFR expression in SCC and ADC for Dako PharmDx and 31G7.

    Techniques: Fluorescence In Situ Hybridization, Amplification

    Immunohistochemical EGFR staining with four different antibodies showing differences in levels of EGFR expression in the same specimen of a squamous cell carcinoma (SSC) (original magnification × 400). (A) Staining intensity with Dako PharmDx 2+, (B) Staining intensity with 31G7 2+, (C) Staining intensity with 2.1E1 3+, (D) Staining intensity with SP84 1+.

    Journal: Diagnostic Pathology

    Article Title: Correlation of EGFR expression, gene copy number and clinicopathological status in NSCLC

    doi: 10.1186/s13000-014-0165-0

    Figure Lengend Snippet: Immunohistochemical EGFR staining with four different antibodies showing differences in levels of EGFR expression in the same specimen of a squamous cell carcinoma (SSC) (original magnification × 400). (A) Staining intensity with Dako PharmDx 2+, (B) Staining intensity with 31G7 2+, (C) Staining intensity with 2.1E1 3+, (D) Staining intensity with SP84 1+.

    Article Snippet: Scoring with method (B) showed a similar EGFR expression in SCC and ADC for Dako PharmDx and 31G7.

    Techniques: Immunohistochemistry, Staining, Expressing

    EGFR (epidermal growth factor receptor) gene status in adrenocortical neoplasms: (a) This image demonstrates strong membrane EGFR expression (3+) in adrenocortical carcinoma, as detected by immunohistochemistry. (b) This image demonstrates an absence of membrane EGFR expression in adrenocortical adenoma, as detected by immunohistochemistry. (c) The cancer cells demonstrate high polysomy on chromosome 7 in adrenocortical carcinoma, as detected by FISH(fluorescence in situ hybridization). (d) The tumor cells display disomy for the EGFR in adrenocortical adenoma, as detected by FISH (Green signals represent the chromosome 7 centromere, and red signals represent the EGFR gene).

    Journal: Diagnostic Pathology

    Article Title: Adrenal cortical neoplasms: a study of clinicopathological features related to epidermal growth factor receptor gene status

    doi: 10.1186/1746-1596-9-19

    Figure Lengend Snippet: EGFR (epidermal growth factor receptor) gene status in adrenocortical neoplasms: (a) This image demonstrates strong membrane EGFR expression (3+) in adrenocortical carcinoma, as detected by immunohistochemistry. (b) This image demonstrates an absence of membrane EGFR expression in adrenocortical adenoma, as detected by immunohistochemistry. (c) The cancer cells demonstrate high polysomy on chromosome 7 in adrenocortical carcinoma, as detected by FISH(fluorescence in situ hybridization). (d) The tumor cells display disomy for the EGFR in adrenocortical adenoma, as detected by FISH (Green signals represent the chromosome 7 centromere, and red signals represent the EGFR gene).

    Article Snippet: Immunohistochemical study EGFR protein expression was evaluated by immunohistochemistry using a mouse anti-human EGFR monoclonal antibody (clone 2-18C9, Pharm Dx kit, Dako North America, Inc., Via Real, Carpinteria, CA, USA), according to the manufacturer’s instructions.

    Techniques: Expressing, Immunohistochemistry, Fluorescence In Situ Hybridization, In Situ Hybridization