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  • 99
    Vector Laboratories vectastain abc reagent
    Vectastain Abc Reagent, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 2984 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Lonza mycoalert mycoplasma detection kit
    Mycoalert Mycoplasma Detection Kit, supplied by Lonza, used in various techniques. Bioz Stars score: 99/100, based on 4660 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi prism 7000 sequence detection system
    Abi Prism 7000 Sequence Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 26856 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare ecl kit
    Ecl Kit, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 20587 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore situ apoptosis detection kit
    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and <t>apoptosis</t> (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P
    Situ Apoptosis Detection Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10129 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher chemiluminescence detection kit
    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and <t>apoptosis</t> (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P
    Chemiluminescence Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5491 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare ecl detection kit
    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and <t>apoptosis</t> (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P
    Ecl Detection Kit, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 7815 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 7815 article reviews
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    Thermo Fisher mirvana qrt pcr mirna detection kit
    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and <t>apoptosis</t> (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P
    Mirvana Qrt Pcr Mirna Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 981 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher ecl detection kit
    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and <t>apoptosis</t> (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P
    Ecl Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2406 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Ventana Medical ultraview universal dab detection kit
    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the <t>ultraView</t> Universal <t>DAB</t> Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.
    Ultraview Universal Dab Detection Kit, supplied by Ventana Medical, used in various techniques. Bioz Stars score: 92/100, based on 2312 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher detection kit
    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the <t>ultraView</t> Universal <t>DAB</t> Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.
    Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2281 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    detection kit - by Bioz Stars, 2021-01
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    Millipore annexin v fitc apoptosis detection kit
    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the <t>ultraView</t> Universal <t>DAB</t> Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.
    Annexin V Fitc Apoptosis Detection Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3130 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Enzo Biochem cyto id autophagy detection kit
    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the <t>ultraView</t> Universal <t>DAB</t> Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.
    Cyto Id Autophagy Detection Kit, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 99/100, based on 1850 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore lookout mycoplasma pcr detection kit
    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the <t>ultraView</t> Universal <t>DAB</t> Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.
    Lookout Mycoplasma Pcr Detection Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa ldh cytotoxicity detection kit
    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the <t>ultraView</t> Universal <t>DAB</t> Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.
    Ldh Cytotoxicity Detection Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 942 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa in situ apoptosis detection kit
    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the <t>ultraView</t> Universal <t>DAB</t> Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.
    In Situ Apoptosis Detection Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1831 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Thermo Fisher mycoplasma contamination
    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the <t>ultraView</t> Universal <t>DAB</t> Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.
    Mycoplasma Contamination, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 669 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam caspase 3
    Protein expression of myocardial tissue in pressure-overloaded heart failure rats. (a) Western blot analysis of eNOS protein expression in myocardial tissue. (b) Quantification of eNOS protein expression in myocardial tissue by densitometry. (c) Western blot analysis of TGF- β 1 protein expression in myocardial tissue. (d) Quantification of TGF- β 1 protein expression in myocardial tissue by densitometry. (e) Western blot analysis of VEGF protein expression in myocardial tissue. (f) Quantification of VEGF protein expression in myocardial tissue by densitometry. (g) Western blot analysis of VEGFR2 protein expression in myocardial tissue. (h) Quantification of VEGFR2 protein expression in myocardial tissue by densitometry. (i) Western blot analysis of <t>caspase-3</t> protein expression in myocardial tissue. (j) Quantification of caspase-3 protein expression in myocardial tissue by densitometry. Data are expressed as mean ± standard deviation (SD). Δ P
    Caspase 3, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 4883 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    N/A
    Biotoxis is a qPCR all in one detection kit that detects B anthracis Y pestis and F tularensis These 3 pathogens are the first potential biological weapon that can cause
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    The BD Pharmingen BrdU In Situ Detection Kit is designed for immunohistochemical staining of BrdU in frozen sections formalin fixed paraffin embedded sections and cultured or isolated cells on slides
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    Image Search Results


    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P

    Article Snippet: For whole mount TUNEL staining and immunofluorescence staining, samples were processed using the ApoTag plus Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7101) according to the manufacturer’s instructions and as previously described .

    Techniques: Activity Assay, Mouse Assay, Two Tailed Test

    TNFR1 signalling drives cell death and lethality of HOIL-1-deficient mice at mid-gestation. a, d , Quantification of genotypes of animals obtained from inter-crosses of Tnf -/- Hoil-1 +/- (a) and Tnfr1 -/- Hoil-1 +/- (d) mice. *: dead embryos. b, Representative images of embryos quantified in (a) at E10.5 and E15.5, *; poor yolk sac. c, Cell death as detected by whole-mount TUNEL staining in yolk sacs at E10.5 (n= 3 embryos/genotype). e, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved Caspase-3, bottom panel) of yolk sacs. Merged image is shown in Scale bar: 50 µm. f, Representative images of embryos at E16.5 ( n =2 for Tnfr1 -/- Hoil-1 +/- and n =4 for Tnfr1 -/- Hoil-1 -/- ). Fig. 1g.

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: TNFR1 signalling drives cell death and lethality of HOIL-1-deficient mice at mid-gestation. a, d , Quantification of genotypes of animals obtained from inter-crosses of Tnf -/- Hoil-1 +/- (a) and Tnfr1 -/- Hoil-1 +/- (d) mice. *: dead embryos. b, Representative images of embryos quantified in (a) at E10.5 and E15.5, *; poor yolk sac. c, Cell death as detected by whole-mount TUNEL staining in yolk sacs at E10.5 (n= 3 embryos/genotype). e, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved Caspase-3, bottom panel) of yolk sacs. Merged image is shown in Scale bar: 50 µm. f, Representative images of embryos at E16.5 ( n =2 for Tnfr1 -/- Hoil-1 +/- and n =4 for Tnfr1 -/- Hoil-1 -/- ). Fig. 1g.

    Article Snippet: For whole mount TUNEL staining and immunofluorescence staining, samples were processed using the ApoTag plus Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7101) according to the manufacturer’s instructions and as previously described .

    Techniques: Mouse Assay, TUNEL Assay, Staining

    HOIL-1-deficient mice die at mid-gestation a, Schematic representation of the Hoil-1 knockout strategy. Solid boxes represent Hoil-1 exons and grey boxes with a star indicate the targeted exons. Boxes with diagonal and horizontal strips represent LoxP and Frt sites, respectively. b, Specificity of gene recombination was assessed by Southern blotting with 5’ and 3’ probes external to the construct in four clones (14B8, 14F6, 20D7 and 21F7). Digest of the DNA with ApaI, followed by hybridisation with the 3’ probe was expected to show a 5700 bp band for the WT allele and a 7700 bp band for the mutant allele. All four clones appeared to have the correct recombination on the 3’ side. Digest of the DNA with SphI and hybridisation with the 5’ probe was expected to show a 4500 bp WT band and a 6200 bp band for the mutated allele. Clones 14B8, 14F6 and 21F7 appeared to be correctly recombined on the 5’ side. Finally, cutting the DNA with ApaI and hybridising with a hygro probe showed a single band in all clones, indicative of a single integration of the construct in all four ES clones. Clones 14B8 and 14F6 were selected for generation of the two Hoil-1 -/- strains. c , PCR analysis of Hoil-1 wild-type, heterozygous and knockout mice. d , Protein levels of HOIL-1, HOIP and SHARPIN in whole embryo lysates ( n = 3 for Hoil-1 +/- and Hoil-1 -/- embryos and n =1 for Hoil-1 +/+ . e , Quantification of genotypes of animals obtained from inter-crossing C20Hoil-1 +/- mice. *indicates dead embryos. f , Representative images of C20Hoil-1 +/- and C20Hoil-1 -/- embryos from E9.5 to E11.5 as quantified in (e). Scale bar: 2 mm. g, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . h , Whole-mount TUNEL staining of embryos at the indicated stages (embryo/genotype n =2 at E10.5, n =8 for Hoil-1 +/- and n =5 for Hoil-1 -/- at E11.5). Scale bar: 2 mm. i, Quantification of genotypes of animals obtained from inter-crossing Hoil-1 fl/wt Tie2-Cre + with Hoil-1 fl/fl Tie2-Cre - mice. *indicates dead embryos. j , Representative images of embryos with conditional deletion of Hoil-1 in Tie2-Cre expressing cells as quantified in (i). Scale bar: 2 mm. *: poorly vascularised yolk sac. Fig. 1c

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: HOIL-1-deficient mice die at mid-gestation a, Schematic representation of the Hoil-1 knockout strategy. Solid boxes represent Hoil-1 exons and grey boxes with a star indicate the targeted exons. Boxes with diagonal and horizontal strips represent LoxP and Frt sites, respectively. b, Specificity of gene recombination was assessed by Southern blotting with 5’ and 3’ probes external to the construct in four clones (14B8, 14F6, 20D7 and 21F7). Digest of the DNA with ApaI, followed by hybridisation with the 3’ probe was expected to show a 5700 bp band for the WT allele and a 7700 bp band for the mutant allele. All four clones appeared to have the correct recombination on the 3’ side. Digest of the DNA with SphI and hybridisation with the 5’ probe was expected to show a 4500 bp WT band and a 6200 bp band for the mutated allele. Clones 14B8, 14F6 and 21F7 appeared to be correctly recombined on the 5’ side. Finally, cutting the DNA with ApaI and hybridising with a hygro probe showed a single band in all clones, indicative of a single integration of the construct in all four ES clones. Clones 14B8 and 14F6 were selected for generation of the two Hoil-1 -/- strains. c , PCR analysis of Hoil-1 wild-type, heterozygous and knockout mice. d , Protein levels of HOIL-1, HOIP and SHARPIN in whole embryo lysates ( n = 3 for Hoil-1 +/- and Hoil-1 -/- embryos and n =1 for Hoil-1 +/+ . e , Quantification of genotypes of animals obtained from inter-crossing C20Hoil-1 +/- mice. *indicates dead embryos. f , Representative images of C20Hoil-1 +/- and C20Hoil-1 -/- embryos from E9.5 to E11.5 as quantified in (e). Scale bar: 2 mm. g, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . h , Whole-mount TUNEL staining of embryos at the indicated stages (embryo/genotype n =2 at E10.5, n =8 for Hoil-1 +/- and n =5 for Hoil-1 -/- at E11.5). Scale bar: 2 mm. i, Quantification of genotypes of animals obtained from inter-crossing Hoil-1 fl/wt Tie2-Cre + with Hoil-1 fl/fl Tie2-Cre - mice. *indicates dead embryos. j , Representative images of embryos with conditional deletion of Hoil-1 in Tie2-Cre expressing cells as quantified in (i). Scale bar: 2 mm. *: poorly vascularised yolk sac. Fig. 1c

    Article Snippet: For whole mount TUNEL staining and immunofluorescence staining, samples were processed using the ApoTag plus Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7101) according to the manufacturer’s instructions and as previously described .

    Techniques: Mouse Assay, Knock-Out, Southern Blot, Construct, Clone Assay, Hybridization, Mutagenesis, Polymerase Chain Reaction, Staining, TUNEL Assay, Expressing

    Individual deletion of mediators of apoptosis or necroptosis does not prevent cell death and lethality at mid-gestation of HOIL-1- or HOIP-deficient embryos. a , Western blot analysis of MLKL expression in the indicated organs derived from control Mlkl -/- mice ( n . b, d, e, f, Representative images of embryos at different stages of gestation (E10.5: n =7 for Ripk3 -/- Hoil-1 +/- and n =5 for Ripk3 -/- Hoil-1 -/- ; E11.5: n =5 for Ripk3 -/- Hoil-1 +/- and n =2 for Ripk3 -/- Hoil-1 -/- ; E12.5: n =9 for Ripk3 -/- Hoil-1 +/- and n =2 for Ripk3 -/- Hoil-1 -/- (b), E10.5: n =16 for Mlkl -/- Hoip +/- and n =6 for Mlkl -/- Hoip -/- ; E11.5: n =8 for Mlkl -/- Hoip +/- and n =6 for Mlkl -/- Hoip -/- ; E12.5: n =10 for Mlkl -/- Hoip +/- and n =5 for Mlkl -/- Hoip -/- (d), E10.5: n =5 for Caspase-8 +/- Hoip +/- and n =4 for Caspase-8 +/- Hoip -/- ; E11.5: n =6 for Caspase-8 +/- Hoip +/- and n =3 for Caspase-8 +/- Hoip -/- ; E12.5: n =3 for Caspase-8 +/- Hoip +/- and n =2 for Caspase-8 +/- Hoip -/- (e), E10.5: n =2 for Caspase-8 +/- Hoil-1 +/- and n =4 for Caspase-8 +/- Hoil-1 -/- ; E11.5: n =2 for Caspase-8 +/- Hoil-1 +/- and n =5 for Caspase-8 +/- Hoil-1 -/- ; E12.5: n =6 for Caspase-8 +/- Hoil-1 +/- and n =3 for Caspase-8 +/- Hoil-1 -/- (f)). *: poor yolk sac vascularisation. Scale bar: 2 mm. c, Representative images of yolk sac vascularisation and cell death at E10.5 as detected by PECAM-1 (red) and cleaved (cl.) Caspase-3 staining (green) (top panel) and whole mount TUNEL staining (bottom panel) ( n =4 per genotype). Scale bar: 50 µm.

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Individual deletion of mediators of apoptosis or necroptosis does not prevent cell death and lethality at mid-gestation of HOIL-1- or HOIP-deficient embryos. a , Western blot analysis of MLKL expression in the indicated organs derived from control Mlkl -/- mice ( n . b, d, e, f, Representative images of embryos at different stages of gestation (E10.5: n =7 for Ripk3 -/- Hoil-1 +/- and n =5 for Ripk3 -/- Hoil-1 -/- ; E11.5: n =5 for Ripk3 -/- Hoil-1 +/- and n =2 for Ripk3 -/- Hoil-1 -/- ; E12.5: n =9 for Ripk3 -/- Hoil-1 +/- and n =2 for Ripk3 -/- Hoil-1 -/- (b), E10.5: n =16 for Mlkl -/- Hoip +/- and n =6 for Mlkl -/- Hoip -/- ; E11.5: n =8 for Mlkl -/- Hoip +/- and n =6 for Mlkl -/- Hoip -/- ; E12.5: n =10 for Mlkl -/- Hoip +/- and n =5 for Mlkl -/- Hoip -/- (d), E10.5: n =5 for Caspase-8 +/- Hoip +/- and n =4 for Caspase-8 +/- Hoip -/- ; E11.5: n =6 for Caspase-8 +/- Hoip +/- and n =3 for Caspase-8 +/- Hoip -/- ; E12.5: n =3 for Caspase-8 +/- Hoip +/- and n =2 for Caspase-8 +/- Hoip -/- (e), E10.5: n =2 for Caspase-8 +/- Hoil-1 +/- and n =4 for Caspase-8 +/- Hoil-1 -/- ; E11.5: n =2 for Caspase-8 +/- Hoil-1 +/- and n =5 for Caspase-8 +/- Hoil-1 -/- ; E12.5: n =6 for Caspase-8 +/- Hoil-1 +/- and n =3 for Caspase-8 +/- Hoil-1 -/- (f)). *: poor yolk sac vascularisation. Scale bar: 2 mm. c, Representative images of yolk sac vascularisation and cell death at E10.5 as detected by PECAM-1 (red) and cleaved (cl.) Caspase-3 staining (green) (top panel) and whole mount TUNEL staining (bottom panel) ( n =4 per genotype). Scale bar: 50 µm.

    Article Snippet: For whole mount TUNEL staining and immunofluorescence staining, samples were processed using the ApoTag plus Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7101) according to the manufacturer’s instructions and as previously described .

    Techniques: Western Blot, Expressing, Derivative Assay, Mouse Assay, Staining, TUNEL Assay

    Combined deletion of MLKL and Caspase-8 promotes survival of LUBAC-deficient mice. a, Quantification of genotypes of animals obtained from inter-crosses of Mlkl -/- Caspase-8 +/- Hoip +/- with Mlkl -/- Caspase-8 -/- Hoip +/- mice. *: dead embryos. b, Representative images of adult mice as quantified in (a). c , Kaplan-Meier plot of mouse survival ( n =6 for Mlkl -/- Caspase-8 -/- Hoip -/- and n =9 for Mlkl -/- Caspase-8 -/- Hoil-1 -/- mice). d, Representative images of H E staining of the indicated organs ( n =3 mice/genotype). Scale bar: 200 µm. e, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) Caspase-3, green) (top panel) at E13.5 and respective quantifications (bottom panel). Mean ± s.e.m ( n =5 for Mlkl -/- Caspase-8 -/- Hoil-1 +/- and Mlkl -/- Caspase-8 -/- Hoil-1 -/- and n =2 for Mlkl -/- Caspase-8 +/- Hoil-1 -/- ) are shown and results were analysed with unpaired two-tailed t -tests comparing Mlkl -/- Caspase-8 -/- Hoil-1 +/- and Mlkl -/- Caspase-8 -/- Hoil-1 -/- embryos. f, Representative images of H E staining of the indicated organs ( n =3 embryos/genotype). Scale bar: 200 µm. g, Epidermal thickness quantification of mice of the indicated genotypes in (f). Mean ± s.e.m values ( n =3 mice/genotype) are shown and results were analysed with unpaired two-tailed t -tests. h, Western blot analysis of lysates from whole E13.5 embryos of the indicated genotypes as well as L929 cells treated or not with TNF plus zVAD-fmk for 2 h as antibody validation ( n .

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Combined deletion of MLKL and Caspase-8 promotes survival of LUBAC-deficient mice. a, Quantification of genotypes of animals obtained from inter-crosses of Mlkl -/- Caspase-8 +/- Hoip +/- with Mlkl -/- Caspase-8 -/- Hoip +/- mice. *: dead embryos. b, Representative images of adult mice as quantified in (a). c , Kaplan-Meier plot of mouse survival ( n =6 for Mlkl -/- Caspase-8 -/- Hoip -/- and n =9 for Mlkl -/- Caspase-8 -/- Hoil-1 -/- mice). d, Representative images of H E staining of the indicated organs ( n =3 mice/genotype). Scale bar: 200 µm. e, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) Caspase-3, green) (top panel) at E13.5 and respective quantifications (bottom panel). Mean ± s.e.m ( n =5 for Mlkl -/- Caspase-8 -/- Hoil-1 +/- and Mlkl -/- Caspase-8 -/- Hoil-1 -/- and n =2 for Mlkl -/- Caspase-8 +/- Hoil-1 -/- ) are shown and results were analysed with unpaired two-tailed t -tests comparing Mlkl -/- Caspase-8 -/- Hoil-1 +/- and Mlkl -/- Caspase-8 -/- Hoil-1 -/- embryos. f, Representative images of H E staining of the indicated organs ( n =3 embryos/genotype). Scale bar: 200 µm. g, Epidermal thickness quantification of mice of the indicated genotypes in (f). Mean ± s.e.m values ( n =3 mice/genotype) are shown and results were analysed with unpaired two-tailed t -tests. h, Western blot analysis of lysates from whole E13.5 embryos of the indicated genotypes as well as L929 cells treated or not with TNF plus zVAD-fmk for 2 h as antibody validation ( n .

    Article Snippet: For whole mount TUNEL staining and immunofluorescence staining, samples were processed using the ApoTag plus Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7101) according to the manufacturer’s instructions and as previously described .

    Techniques: Mouse Assay, Staining, Two Tailed Test, Western Blot

    Combined deletion of RIPK3 and Caspase-8 prevents cell death but not embryonic lethality at late gestation that is caused by the loss of HOIL-1. a , Quantification of genotypes of animals obtained from inter-crosses of Ripk3 -/- Caspase-8 +/- Hoil-1 +/- with Ripk3 -/- Caspase-8 -/- Hoil-1 +/- mice (left panel) or Ripk3 -/- Caspase-8 -/- Hoil-1 +/- mice (right panel). b, Health status of Ripk3 -/- Caspase-8 +/- Hoil-1 -/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- embryos at different developmental stages. c , Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved (cl.) Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . Scale bar: 50 µm. d, Cell death as detected by whole-mount TUNEL staining in yolk sacs at E14.5 (left panel) and respective quantification (right panel). Mean ± s.e.m. ( n =3 embryos/genotype) and P values from one-way ANOVA are reported. e, f Representative images (e) and quantification (f) of cell death in different organs as detected by TUNEL staining at E13.5 ( n =3 embryos/genotype) and E14.5 ( n =5 for Ripk3 -/- Caspase-8 -/- Hoil-1 +/- , n=2 for Ripk3 -/- Caspase-8 -/- Hoil-1 -/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- lung and liver and n =3 Ripk3 -/- Caspase-8 -/- Hoil-1 -/- heart). Scale bar: 50 µm (e). Mean ± s.e.m. values are shown (f). g, Cell death was analysed by PI staining in MEFs stimulated or not with the indicated ligands for 24 h. Mean ± s.e.m. ( n =3 independent experiments) and P values from two-way ANOVA are reported. h, Representative images of H E staining on E13.5 whole-embryo paraffin embedded sections ( n =3 for Ripk3 -/- Caspase-8 -/- Hoil-1 +/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- and n =2 for Ripk3 -/- Caspase-8 +/- Hoil-1 -/- ). *: pericardial effusion, arrows; congested vessels H, heart; L, lung; Li, liver. Scale bar: 200 µm. i, Representative images of micro-focus CT scan images of whole E13.5 embryos ( n =3 embryos/genotype). *: pericardial effusion Fig. 3f

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Combined deletion of RIPK3 and Caspase-8 prevents cell death but not embryonic lethality at late gestation that is caused by the loss of HOIL-1. a , Quantification of genotypes of animals obtained from inter-crosses of Ripk3 -/- Caspase-8 +/- Hoil-1 +/- with Ripk3 -/- Caspase-8 -/- Hoil-1 +/- mice (left panel) or Ripk3 -/- Caspase-8 -/- Hoil-1 +/- mice (right panel). b, Health status of Ripk3 -/- Caspase-8 +/- Hoil-1 -/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- embryos at different developmental stages. c , Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved (cl.) Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . Scale bar: 50 µm. d, Cell death as detected by whole-mount TUNEL staining in yolk sacs at E14.5 (left panel) and respective quantification (right panel). Mean ± s.e.m. ( n =3 embryos/genotype) and P values from one-way ANOVA are reported. e, f Representative images (e) and quantification (f) of cell death in different organs as detected by TUNEL staining at E13.5 ( n =3 embryos/genotype) and E14.5 ( n =5 for Ripk3 -/- Caspase-8 -/- Hoil-1 +/- , n=2 for Ripk3 -/- Caspase-8 -/- Hoil-1 -/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- lung and liver and n =3 Ripk3 -/- Caspase-8 -/- Hoil-1 -/- heart). Scale bar: 50 µm (e). Mean ± s.e.m. values are shown (f). g, Cell death was analysed by PI staining in MEFs stimulated or not with the indicated ligands for 24 h. Mean ± s.e.m. ( n =3 independent experiments) and P values from two-way ANOVA are reported. h, Representative images of H E staining on E13.5 whole-embryo paraffin embedded sections ( n =3 for Ripk3 -/- Caspase-8 -/- Hoil-1 +/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- and n =2 for Ripk3 -/- Caspase-8 +/- Hoil-1 -/- ). *: pericardial effusion, arrows; congested vessels H, heart; L, lung; Li, liver. Scale bar: 200 µm. i, Representative images of micro-focus CT scan images of whole E13.5 embryos ( n =3 embryos/genotype). *: pericardial effusion Fig. 3f

    Article Snippet: For whole mount TUNEL staining and immunofluorescence staining, samples were processed using the ApoTag plus Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7101) according to the manufacturer’s instructions and as previously described .

    Techniques: Mouse Assay, Staining, TUNEL Assay, Computed Tomography

    Ablation of the kinase activity of RIPK1 in HOIL-1- or HOIP-deficient embryos prevents cell death and lethality at mid-gestation but not at late gestation. a, b, Quantification of genotypes of animals obtained after inter-crossing Ripk1 K45A Hoil-1 +/- (a) and Ripk1 K45A Hoip +/- (b) mice. *indicates dead embryos. c, Representative images of embryos quantified in (b) *; poor yolk sac vascularisation. Scale bar: 2 mm. d, Whole-mount TUNEL staining of embryos ( n =2 embryos). Scale bar: 2 mm. e, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved (cl.) Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . f, g, Representative images of cell death in different organs (f) and quantification (g) as detected by TUNEL staining at E14.5 ( n =3 embryos/genotype). Scale bar: 50 μm (f). Mean ± s.e.m. ( n =3 embryos/genotype) and P values from one-way ANOVA are reported (g). h, Representative images of H E staining on whole-embryo paraffin sections ( n =3 embryos/genotype). *; pericardial effusion, n, necrotic area. H, heart; L, lung; Li, liver. Scale bar: 200 µm. i, Cell death was analysed by PI staining in MEFs stimulated or not with TNF for 24 h plus the indicated cell death inhibitors. Mean ± s.e.m. ( n =3 independent experiments) and P values from two-way ANOVA are reported. Fig. 3b

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Ablation of the kinase activity of RIPK1 in HOIL-1- or HOIP-deficient embryos prevents cell death and lethality at mid-gestation but not at late gestation. a, b, Quantification of genotypes of animals obtained after inter-crossing Ripk1 K45A Hoil-1 +/- (a) and Ripk1 K45A Hoip +/- (b) mice. *indicates dead embryos. c, Representative images of embryos quantified in (b) *; poor yolk sac vascularisation. Scale bar: 2 mm. d, Whole-mount TUNEL staining of embryos ( n =2 embryos). Scale bar: 2 mm. e, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved (cl.) Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . f, g, Representative images of cell death in different organs (f) and quantification (g) as detected by TUNEL staining at E14.5 ( n =3 embryos/genotype). Scale bar: 50 μm (f). Mean ± s.e.m. ( n =3 embryos/genotype) and P values from one-way ANOVA are reported (g). h, Representative images of H E staining on whole-embryo paraffin sections ( n =3 embryos/genotype). *; pericardial effusion, n, necrotic area. H, heart; L, lung; Li, liver. Scale bar: 200 µm. i, Cell death was analysed by PI staining in MEFs stimulated or not with TNF for 24 h plus the indicated cell death inhibitors. Mean ± s.e.m. ( n =3 independent experiments) and P values from two-way ANOVA are reported. Fig. 3b

    Article Snippet: For whole mount TUNEL staining and immunofluorescence staining, samples were processed using the ApoTag plus Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7101) according to the manufacturer’s instructions and as previously described .

    Techniques: Activity Assay, Mouse Assay, TUNEL Assay, Staining

    HOIL-1 deficiency causes embryonic lethality at mid-gestation due to TNFR1-mediated endothelial cell death a, Mendelian frequencies obtained from inter-crossing Hoil-1 +/- mice, *: dead embryos. b, Representative images of embryos from E9.5 to E11.5 quantified in (a), *: poor yolk sac vascularisation. Scale bar: 2 mm. c, Representative images of yolk sac vascularisation (PECAM-1, red) and cell death (cleaved (cl.) Caspase-3 staining, green) at E10.5 (top panel) ( n =4 yolk-sacs/genotype), and whole-mount TUNEL staining (bottom, panel) ( n =2 yolk-sacs/genotype). Scale bar: 50 µm. d, g, Quantification of branching points (g) and cleaved Caspase-3 positive cells (g). Mean ± s.e.m. values and P values from unpaired two-tailed t -tests are shown. e, Representative images of embryos at E10.5 ( n =14 Hoil-1 fl/wt Tie2-Cre+ and n =7 Hoil-1 fl/fl Tie2-Cre+ embryos, top panel). *: poor yolk sac vascularisation. Scale bar: 2 mm. Yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved Caspase-3, green) (middle panel). Scale bar: 50 µm. Yolk sac whole-mount TUNEL staining ( n =6 Hoil-1 fl/wt Tie2-Cre+ and n =2 Hoil-1 fl/fl Tie2-Cre+ yolk-sacs/genotype , bottom panel). f , Representative images of embryos at E15.5 (top panel, n =6 Tnfr1 -/- Hoil-1 -/- and n =19 Tnfr1 -/- Hoil-1 +/- embryos), scale bar: 2 mm, and yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved Caspase-3, green) (bottom panel), Scale bar: 50 µm. h, Representative images of H E staining on whole-embryo paraffin sections ( n =3 embryos/genotype). *: pericardial effusion, arrows; congested vessels. H, heart; L, lung; Li, liver. Scale bar: 50 µm.

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: HOIL-1 deficiency causes embryonic lethality at mid-gestation due to TNFR1-mediated endothelial cell death a, Mendelian frequencies obtained from inter-crossing Hoil-1 +/- mice, *: dead embryos. b, Representative images of embryos from E9.5 to E11.5 quantified in (a), *: poor yolk sac vascularisation. Scale bar: 2 mm. c, Representative images of yolk sac vascularisation (PECAM-1, red) and cell death (cleaved (cl.) Caspase-3 staining, green) at E10.5 (top panel) ( n =4 yolk-sacs/genotype), and whole-mount TUNEL staining (bottom, panel) ( n =2 yolk-sacs/genotype). Scale bar: 50 µm. d, g, Quantification of branching points (g) and cleaved Caspase-3 positive cells (g). Mean ± s.e.m. values and P values from unpaired two-tailed t -tests are shown. e, Representative images of embryos at E10.5 ( n =14 Hoil-1 fl/wt Tie2-Cre+ and n =7 Hoil-1 fl/fl Tie2-Cre+ embryos, top panel). *: poor yolk sac vascularisation. Scale bar: 2 mm. Yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved Caspase-3, green) (middle panel). Scale bar: 50 µm. Yolk sac whole-mount TUNEL staining ( n =6 Hoil-1 fl/wt Tie2-Cre+ and n =2 Hoil-1 fl/fl Tie2-Cre+ yolk-sacs/genotype , bottom panel). f , Representative images of embryos at E15.5 (top panel, n =6 Tnfr1 -/- Hoil-1 -/- and n =19 Tnfr1 -/- Hoil-1 +/- embryos), scale bar: 2 mm, and yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved Caspase-3, green) (bottom panel), Scale bar: 50 µm. h, Representative images of H E staining on whole-embryo paraffin sections ( n =3 embryos/genotype). *: pericardial effusion, arrows; congested vessels. H, heart; L, lung; Li, liver. Scale bar: 50 µm.

    Article Snippet: For whole mount TUNEL staining and immunofluorescence staining, samples were processed using the ApoTag plus Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7101) according to the manufacturer’s instructions and as previously described .

    Techniques: Mouse Assay, Staining, TUNEL Assay, Two Tailed Test

    (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the ultraView Universal DAB Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.

    Journal: International Journal of Molecular Medicine

    Article Title: Human umbilical cord blood mononuclear cells and chorionic plate-derived mesenchymal stem cells promote axon survival in a rat model of optic nerve crush injury

    doi: 10.3892/ijmm.2016.2532

    Figure Lengend Snippet: (A) Sequential histological findings using hematoxylin and eosin staining of optic nerve samples from a rat model of optic nerve compression after injection of umbilical cord blood mononuclear cells (CB-MNCs) or chorionic plate-derived MSCs (CP-MSCs). At 2 weeks post-CB-MNC injection, subacute inflammation was observed in the optic nerve around the compression area (arrow), compared with the controls. At 2 weeks post-CP-MSC injection, subacute inflammation was observed in the optic nerve around the compression line (arrow), compared with the control. (B) Immunohistochemical findings in the optic nerve after injection with CB-MNCs or CP-MSCs using 5- µ m-thick sections cut from formalin-fixed, paraffin wax-embedded tissue and the anti-human nuclei and chromosomes, histone H1 protein antibody (mouse monoclonal, clone 1415-1, 1:200 dilution). The BenchMark XT was used as the visualization system together with heat-induced epitope retrieval (CC1 solution) and the ultraView Universal DAB Detection kit. At 2 weeks post-CB-MNC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). Similarly, at 2 weeks post-CP-MSC injection, the presence of surviving human MNCs was hardly identified immunohistologically due to strong cross reactivity (dark brown nuclei around the compression area (arrow) in the optic nerve compared with the control). All images are ×400 magnification. Control, non-transplanted group; CB-MNC, CB-MNC-transplanted group; CP-MSC, CP-MSC-transplanted group.

    Article Snippet: The visualization system used was the BenchMark XT with heat-induced epitope retrieval (CC1 solution) and the ultraView Universal DAB Detection kit (all from Ventana, Tucson, AZ, USA).

    Techniques: Staining, Injection, Derivative Assay, Immunohistochemistry

    Protein expression of myocardial tissue in pressure-overloaded heart failure rats. (a) Western blot analysis of eNOS protein expression in myocardial tissue. (b) Quantification of eNOS protein expression in myocardial tissue by densitometry. (c) Western blot analysis of TGF- β 1 protein expression in myocardial tissue. (d) Quantification of TGF- β 1 protein expression in myocardial tissue by densitometry. (e) Western blot analysis of VEGF protein expression in myocardial tissue. (f) Quantification of VEGF protein expression in myocardial tissue by densitometry. (g) Western blot analysis of VEGFR2 protein expression in myocardial tissue. (h) Quantification of VEGFR2 protein expression in myocardial tissue by densitometry. (i) Western blot analysis of caspase-3 protein expression in myocardial tissue. (j) Quantification of caspase-3 protein expression in myocardial tissue by densitometry. Data are expressed as mean ± standard deviation (SD). Δ P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Luhong Formula Has a Cardioprotective Effect on Left Ventricular Remodeling in Pressure-Overloaded Rats

    doi: 10.1155/2020/4095967

    Figure Lengend Snippet: Protein expression of myocardial tissue in pressure-overloaded heart failure rats. (a) Western blot analysis of eNOS protein expression in myocardial tissue. (b) Quantification of eNOS protein expression in myocardial tissue by densitometry. (c) Western blot analysis of TGF- β 1 protein expression in myocardial tissue. (d) Quantification of TGF- β 1 protein expression in myocardial tissue by densitometry. (e) Western blot analysis of VEGF protein expression in myocardial tissue. (f) Quantification of VEGF protein expression in myocardial tissue by densitometry. (g) Western blot analysis of VEGFR2 protein expression in myocardial tissue. (h) Quantification of VEGFR2 protein expression in myocardial tissue by densitometry. (i) Western blot analysis of caspase-3 protein expression in myocardial tissue. (j) Quantification of caspase-3 protein expression in myocardial tissue by densitometry. Data are expressed as mean ± standard deviation (SD). Δ P

    Article Snippet: The blots were reacted with antibodies to eNOS (1 : 500, Abcam, Ab50010), TGF-β1 (1 : 800, Abcam, Ab92486), caspase-3 (1 : 500, CST, 14220T), VEGF(1 : 1000, Abcam, Ab46154), and VEGFR2 (1 : 1000, Abcam, Ab11939) overnight at 4°C and then with secondary antibody at 37°C for 1 hour.

    Techniques: Expressing, Western Blot, Standard Deviation