plasmid cag human cd63 gfp  (Lonza)


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    Lonza plasmid cag human cd63 gfp
    Human <t>CD63-GFP</t> localization and extracellular vesicle (EV) analysis in the primary fibroblast cells obtained from the caudal vertebrae of Tg rats. ( a ) Localization of human CD63-GFP in the cultured Tg rat cells. Immunostaining indicated the co-localization of GFP with human CD63 (upper panels) and with rat CD63-positive signals (lower panels) around nuclei (blue). Scale bars = 50 μm. ( b ) Size distribution of the EVs isolated from the conditioned medium of Wt and Tg rat cells was determined using a NanoSight system. ( c , d ) The relationship between ceramide and the secretion of EVs. The intracellular rat CD63-positive signals and GFP signals were increased after treatment with 10 μM GW4869, a neutral sphingomyelinase (nSMase) inhibitor, for 24 hours ( c ). Western blotting showed a GW4869-dependent decrease of EV markers (rat CD63 and flotillin-1) and human CD63-GFP in the isolated EVs ( d ; left). However, the expression levels of rat CD63 and human CD63-GFP in the cell lysates were not changed by GW4869 ( d ; right). β-actin was used as a loading control. Both generation and protein composition of the EVs did not show an apparent change by CD63-GFP overexpression ( Supplementary Fig. 5 ).
    Plasmid Cag Human Cd63 Gfp, supplied by Lonza, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid cag human cd63 gfp/product/Lonza
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plasmid cag human cd63 gfp - by Bioz Stars, 2020-09
    90/100 stars

    Related Products / Commonly Used Together

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    Images

    1) Product Images from "Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids"

    Article Title: Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids

    Journal: Scientific Reports

    doi: 10.1038/srep31172

    Human CD63-GFP localization and extracellular vesicle (EV) analysis in the primary fibroblast cells obtained from the caudal vertebrae of Tg rats. ( a ) Localization of human CD63-GFP in the cultured Tg rat cells. Immunostaining indicated the co-localization of GFP with human CD63 (upper panels) and with rat CD63-positive signals (lower panels) around nuclei (blue). Scale bars = 50 μm. ( b ) Size distribution of the EVs isolated from the conditioned medium of Wt and Tg rat cells was determined using a NanoSight system. ( c , d ) The relationship between ceramide and the secretion of EVs. The intracellular rat CD63-positive signals and GFP signals were increased after treatment with 10 μM GW4869, a neutral sphingomyelinase (nSMase) inhibitor, for 24 hours ( c ). Western blotting showed a GW4869-dependent decrease of EV markers (rat CD63 and flotillin-1) and human CD63-GFP in the isolated EVs ( d ; left). However, the expression levels of rat CD63 and human CD63-GFP in the cell lysates were not changed by GW4869 ( d ; right). β-actin was used as a loading control. Both generation and protein composition of the EVs did not show an apparent change by CD63-GFP overexpression ( Supplementary Fig. 5 ).
    Figure Legend Snippet: Human CD63-GFP localization and extracellular vesicle (EV) analysis in the primary fibroblast cells obtained from the caudal vertebrae of Tg rats. ( a ) Localization of human CD63-GFP in the cultured Tg rat cells. Immunostaining indicated the co-localization of GFP with human CD63 (upper panels) and with rat CD63-positive signals (lower panels) around nuclei (blue). Scale bars = 50 μm. ( b ) Size distribution of the EVs isolated from the conditioned medium of Wt and Tg rat cells was determined using a NanoSight system. ( c , d ) The relationship between ceramide and the secretion of EVs. The intracellular rat CD63-positive signals and GFP signals were increased after treatment with 10 μM GW4869, a neutral sphingomyelinase (nSMase) inhibitor, for 24 hours ( c ). Western blotting showed a GW4869-dependent decrease of EV markers (rat CD63 and flotillin-1) and human CD63-GFP in the isolated EVs ( d ; left). However, the expression levels of rat CD63 and human CD63-GFP in the cell lysates were not changed by GW4869 ( d ; right). β-actin was used as a loading control. Both generation and protein composition of the EVs did not show an apparent change by CD63-GFP overexpression ( Supplementary Fig. 5 ).

    Techniques Used: Cell Culture, Immunostaining, Isolation, Western Blot, Expressing, Over Expression

    Human CD63-GFP expression analysis in Tg rats (Wister-esTgN(CAG/CD63-GFP)3NCCRI). ( a ) Pictures of main organs from Tg offspring (i–xiii: bright field, i’–xiii’: GFP, and xiii”: merged). GFP-negative (i and i’) and GFP-positive (ii and ii’) offspring were littermate. The heart, kidneys and stomach showed especially high fluorescent signals (xiii”). ( b ) Western blotting for endogenous rat CD63 and exogenous human CD63 in tissue lysates from GFP-negative (GFP−) and GFP-positive (GFP+) offspring. Ctx: cortex, cbl: cerebellum, and hip: hippocampus.
    Figure Legend Snippet: Human CD63-GFP expression analysis in Tg rats (Wister-esTgN(CAG/CD63-GFP)3NCCRI). ( a ) Pictures of main organs from Tg offspring (i–xiii: bright field, i’–xiii’: GFP, and xiii”: merged). GFP-negative (i and i’) and GFP-positive (ii and ii’) offspring were littermate. The heart, kidneys and stomach showed especially high fluorescent signals (xiii”). ( b ) Western blotting for endogenous rat CD63 and exogenous human CD63 in tissue lysates from GFP-negative (GFP−) and GFP-positive (GFP+) offspring. Ctx: cortex, cbl: cerebellum, and hip: hippocampus.

    Techniques Used: Expressing, Western Blot

    Analysis of the EVs labelled with human CD63-GFP in the three body fluids. ( a , b ) Western blotting analysis of the EVs isolated from serum ( a ), breast milk and AF ( b ) of Wt and Tg rats for flotillin-1, rat CD63, human CD63 and copGFP. AF samples were collected from pregnant Tg rats at E16–17 after mating with Wt males. GFP−: GFP-negative foetuses. GFP+: GFP-positive foetuses. ( c ) Immunoelectron microscopy images of serum-derived EVs from Wt and Tg rats using anti-human CD63 antibody (10 nm gold particles). Scale bars = 200 nm.
    Figure Legend Snippet: Analysis of the EVs labelled with human CD63-GFP in the three body fluids. ( a , b ) Western blotting analysis of the EVs isolated from serum ( a ), breast milk and AF ( b ) of Wt and Tg rats for flotillin-1, rat CD63, human CD63 and copGFP. AF samples were collected from pregnant Tg rats at E16–17 after mating with Wt males. GFP−: GFP-negative foetuses. GFP+: GFP-positive foetuses. ( c ) Immunoelectron microscopy images of serum-derived EVs from Wt and Tg rats using anti-human CD63 antibody (10 nm gold particles). Scale bars = 200 nm.

    Techniques Used: Western Blot, Isolation, Immuno-Electron Microscopy, Derivative Assay

    Generation of CAG/human CD63-GFP transgenic (Tg) rats. ( a ) Structure of the transgene construction. The transgene was constructed using human CD63-copGFP under control of the CAG promoter. ( b ) Image of rat embryonic stem cells (rESCs) transfected with the CAG/human CD63-GFP gene. The cultured rESCs expressed GFP. ( c ) Blastocysts after microinjection of the transfected rESCs. The arrow indicates rESC adherence to the inner cell mass (ICM). BF: bright field. Scale bars = 100 μm. ( d ) Adult female chimaeric rat from Wister-derived rESC (white-coated) injection into LEA blastocysts (brown-coated). White patches were present in the face (arrowhead). Two Tg offspring (white-coated) from mating a female chimaeric rat with a Wistar wild type (Wt) male (arrows). ( e ) Genotyping by PCR analysis of the extracted DNA from ear snips of the offspring. B: brown coat colour, W: white coat colour, V: CAG/human CD63-GFP vector, and M: size marker.
    Figure Legend Snippet: Generation of CAG/human CD63-GFP transgenic (Tg) rats. ( a ) Structure of the transgene construction. The transgene was constructed using human CD63-copGFP under control of the CAG promoter. ( b ) Image of rat embryonic stem cells (rESCs) transfected with the CAG/human CD63-GFP gene. The cultured rESCs expressed GFP. ( c ) Blastocysts after microinjection of the transfected rESCs. The arrow indicates rESC adherence to the inner cell mass (ICM). BF: bright field. Scale bars = 100 μm. ( d ) Adult female chimaeric rat from Wister-derived rESC (white-coated) injection into LEA blastocysts (brown-coated). White patches were present in the face (arrowhead). Two Tg offspring (white-coated) from mating a female chimaeric rat with a Wistar wild type (Wt) male (arrows). ( e ) Genotyping by PCR analysis of the extracted DNA from ear snips of the offspring. B: brown coat colour, W: white coat colour, V: CAG/human CD63-GFP vector, and M: size marker.

    Techniques Used: Transgenic Assay, Construct, Transfection, Cell Culture, Derivative Assay, Injection, Polymerase Chain Reaction, Plasmid Preparation, Marker

    Related Articles

    Transfection:

    Article Title: Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids
    Article Snippet: .. The plasmid CAG/human CD63-GFP was linearized by SalI and was then transfected into Wistar rESCs using a Mouse ES Cell Nucleofector Kit (Lonza, Basel, Swiss) and an Amaxa Nucleofector (A-013 program) as described previously . pCAG/human CD63-GFP contained a neomycin/kanamycin selection cassette. .. The rESCs were seeded onto mitomycin-C-treated neomycin-resistant MEFs (Millipore, MA, USA) in YPAC medium with 2% Matrigel (Becton Dickinson, NJ, USA).

    Selection:

    Article Title: Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids
    Article Snippet: .. The plasmid CAG/human CD63-GFP was linearized by SalI and was then transfected into Wistar rESCs using a Mouse ES Cell Nucleofector Kit (Lonza, Basel, Swiss) and an Amaxa Nucleofector (A-013 program) as described previously . pCAG/human CD63-GFP contained a neomycin/kanamycin selection cassette. .. The rESCs were seeded onto mitomycin-C-treated neomycin-resistant MEFs (Millipore, MA, USA) in YPAC medium with 2% Matrigel (Becton Dickinson, NJ, USA).

    Plasmid Preparation:

    Article Title: Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids
    Article Snippet: .. The plasmid CAG/human CD63-GFP was linearized by SalI and was then transfected into Wistar rESCs using a Mouse ES Cell Nucleofector Kit (Lonza, Basel, Swiss) and an Amaxa Nucleofector (A-013 program) as described previously . pCAG/human CD63-GFP contained a neomycin/kanamycin selection cassette. .. The rESCs were seeded onto mitomycin-C-treated neomycin-resistant MEFs (Millipore, MA, USA) in YPAC medium with 2% Matrigel (Becton Dickinson, NJ, USA).

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    Lonza plasmid cag human cd63 gfp
    Human <t>CD63-GFP</t> localization and extracellular vesicle (EV) analysis in the primary fibroblast cells obtained from the caudal vertebrae of Tg rats. ( a ) Localization of human CD63-GFP in the cultured Tg rat cells. Immunostaining indicated the co-localization of GFP with human CD63 (upper panels) and with rat CD63-positive signals (lower panels) around nuclei (blue). Scale bars = 50 μm. ( b ) Size distribution of the EVs isolated from the conditioned medium of Wt and Tg rat cells was determined using a NanoSight system. ( c , d ) The relationship between ceramide and the secretion of EVs. The intracellular rat CD63-positive signals and GFP signals were increased after treatment with 10 μM GW4869, a neutral sphingomyelinase (nSMase) inhibitor, for 24 hours ( c ). Western blotting showed a GW4869-dependent decrease of EV markers (rat CD63 and flotillin-1) and human CD63-GFP in the isolated EVs ( d ; left). However, the expression levels of rat CD63 and human CD63-GFP in the cell lysates were not changed by GW4869 ( d ; right). β-actin was used as a loading control. Both generation and protein composition of the EVs did not show an apparent change by CD63-GFP overexpression ( Supplementary Fig. 5 ).
    Plasmid Cag Human Cd63 Gfp, supplied by Lonza, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid cag human cd63 gfp/product/Lonza
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plasmid cag human cd63 gfp - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    Image Search Results


    Human CD63-GFP localization and extracellular vesicle (EV) analysis in the primary fibroblast cells obtained from the caudal vertebrae of Tg rats. ( a ) Localization of human CD63-GFP in the cultured Tg rat cells. Immunostaining indicated the co-localization of GFP with human CD63 (upper panels) and with rat CD63-positive signals (lower panels) around nuclei (blue). Scale bars = 50 μm. ( b ) Size distribution of the EVs isolated from the conditioned medium of Wt and Tg rat cells was determined using a NanoSight system. ( c , d ) The relationship between ceramide and the secretion of EVs. The intracellular rat CD63-positive signals and GFP signals were increased after treatment with 10 μM GW4869, a neutral sphingomyelinase (nSMase) inhibitor, for 24 hours ( c ). Western blotting showed a GW4869-dependent decrease of EV markers (rat CD63 and flotillin-1) and human CD63-GFP in the isolated EVs ( d ; left). However, the expression levels of rat CD63 and human CD63-GFP in the cell lysates were not changed by GW4869 ( d ; right). β-actin was used as a loading control. Both generation and protein composition of the EVs did not show an apparent change by CD63-GFP overexpression ( Supplementary Fig. 5 ).

    Journal: Scientific Reports

    Article Title: Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids

    doi: 10.1038/srep31172

    Figure Lengend Snippet: Human CD63-GFP localization and extracellular vesicle (EV) analysis in the primary fibroblast cells obtained from the caudal vertebrae of Tg rats. ( a ) Localization of human CD63-GFP in the cultured Tg rat cells. Immunostaining indicated the co-localization of GFP with human CD63 (upper panels) and with rat CD63-positive signals (lower panels) around nuclei (blue). Scale bars = 50 μm. ( b ) Size distribution of the EVs isolated from the conditioned medium of Wt and Tg rat cells was determined using a NanoSight system. ( c , d ) The relationship between ceramide and the secretion of EVs. The intracellular rat CD63-positive signals and GFP signals were increased after treatment with 10 μM GW4869, a neutral sphingomyelinase (nSMase) inhibitor, for 24 hours ( c ). Western blotting showed a GW4869-dependent decrease of EV markers (rat CD63 and flotillin-1) and human CD63-GFP in the isolated EVs ( d ; left). However, the expression levels of rat CD63 and human CD63-GFP in the cell lysates were not changed by GW4869 ( d ; right). β-actin was used as a loading control. Both generation and protein composition of the EVs did not show an apparent change by CD63-GFP overexpression ( Supplementary Fig. 5 ).

    Article Snippet: The plasmid CAG/human CD63-GFP was linearized by SalI and was then transfected into Wistar rESCs using a Mouse ES Cell Nucleofector Kit (Lonza, Basel, Swiss) and an Amaxa Nucleofector (A-013 program) as described previously . pCAG/human CD63-GFP contained a neomycin/kanamycin selection cassette.

    Techniques: Cell Culture, Immunostaining, Isolation, Western Blot, Expressing, Over Expression

    Human CD63-GFP expression analysis in Tg rats (Wister-esTgN(CAG/CD63-GFP)3NCCRI). ( a ) Pictures of main organs from Tg offspring (i–xiii: bright field, i’–xiii’: GFP, and xiii”: merged). GFP-negative (i and i’) and GFP-positive (ii and ii’) offspring were littermate. The heart, kidneys and stomach showed especially high fluorescent signals (xiii”). ( b ) Western blotting for endogenous rat CD63 and exogenous human CD63 in tissue lysates from GFP-negative (GFP−) and GFP-positive (GFP+) offspring. Ctx: cortex, cbl: cerebellum, and hip: hippocampus.

    Journal: Scientific Reports

    Article Title: Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids

    doi: 10.1038/srep31172

    Figure Lengend Snippet: Human CD63-GFP expression analysis in Tg rats (Wister-esTgN(CAG/CD63-GFP)3NCCRI). ( a ) Pictures of main organs from Tg offspring (i–xiii: bright field, i’–xiii’: GFP, and xiii”: merged). GFP-negative (i and i’) and GFP-positive (ii and ii’) offspring were littermate. The heart, kidneys and stomach showed especially high fluorescent signals (xiii”). ( b ) Western blotting for endogenous rat CD63 and exogenous human CD63 in tissue lysates from GFP-negative (GFP−) and GFP-positive (GFP+) offspring. Ctx: cortex, cbl: cerebellum, and hip: hippocampus.

    Article Snippet: The plasmid CAG/human CD63-GFP was linearized by SalI and was then transfected into Wistar rESCs using a Mouse ES Cell Nucleofector Kit (Lonza, Basel, Swiss) and an Amaxa Nucleofector (A-013 program) as described previously . pCAG/human CD63-GFP contained a neomycin/kanamycin selection cassette.

    Techniques: Expressing, Western Blot

    Analysis of the EVs labelled with human CD63-GFP in the three body fluids. ( a , b ) Western blotting analysis of the EVs isolated from serum ( a ), breast milk and AF ( b ) of Wt and Tg rats for flotillin-1, rat CD63, human CD63 and copGFP. AF samples were collected from pregnant Tg rats at E16–17 after mating with Wt males. GFP−: GFP-negative foetuses. GFP+: GFP-positive foetuses. ( c ) Immunoelectron microscopy images of serum-derived EVs from Wt and Tg rats using anti-human CD63 antibody (10 nm gold particles). Scale bars = 200 nm.

    Journal: Scientific Reports

    Article Title: Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids

    doi: 10.1038/srep31172

    Figure Lengend Snippet: Analysis of the EVs labelled with human CD63-GFP in the three body fluids. ( a , b ) Western blotting analysis of the EVs isolated from serum ( a ), breast milk and AF ( b ) of Wt and Tg rats for flotillin-1, rat CD63, human CD63 and copGFP. AF samples were collected from pregnant Tg rats at E16–17 after mating with Wt males. GFP−: GFP-negative foetuses. GFP+: GFP-positive foetuses. ( c ) Immunoelectron microscopy images of serum-derived EVs from Wt and Tg rats using anti-human CD63 antibody (10 nm gold particles). Scale bars = 200 nm.

    Article Snippet: The plasmid CAG/human CD63-GFP was linearized by SalI and was then transfected into Wistar rESCs using a Mouse ES Cell Nucleofector Kit (Lonza, Basel, Swiss) and an Amaxa Nucleofector (A-013 program) as described previously . pCAG/human CD63-GFP contained a neomycin/kanamycin selection cassette.

    Techniques: Western Blot, Isolation, Immuno-Electron Microscopy, Derivative Assay

    Generation of CAG/human CD63-GFP transgenic (Tg) rats. ( a ) Structure of the transgene construction. The transgene was constructed using human CD63-copGFP under control of the CAG promoter. ( b ) Image of rat embryonic stem cells (rESCs) transfected with the CAG/human CD63-GFP gene. The cultured rESCs expressed GFP. ( c ) Blastocysts after microinjection of the transfected rESCs. The arrow indicates rESC adherence to the inner cell mass (ICM). BF: bright field. Scale bars = 100 μm. ( d ) Adult female chimaeric rat from Wister-derived rESC (white-coated) injection into LEA blastocysts (brown-coated). White patches were present in the face (arrowhead). Two Tg offspring (white-coated) from mating a female chimaeric rat with a Wistar wild type (Wt) male (arrows). ( e ) Genotyping by PCR analysis of the extracted DNA from ear snips of the offspring. B: brown coat colour, W: white coat colour, V: CAG/human CD63-GFP vector, and M: size marker.

    Journal: Scientific Reports

    Article Title: Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids

    doi: 10.1038/srep31172

    Figure Lengend Snippet: Generation of CAG/human CD63-GFP transgenic (Tg) rats. ( a ) Structure of the transgene construction. The transgene was constructed using human CD63-copGFP under control of the CAG promoter. ( b ) Image of rat embryonic stem cells (rESCs) transfected with the CAG/human CD63-GFP gene. The cultured rESCs expressed GFP. ( c ) Blastocysts after microinjection of the transfected rESCs. The arrow indicates rESC adherence to the inner cell mass (ICM). BF: bright field. Scale bars = 100 μm. ( d ) Adult female chimaeric rat from Wister-derived rESC (white-coated) injection into LEA blastocysts (brown-coated). White patches were present in the face (arrowhead). Two Tg offspring (white-coated) from mating a female chimaeric rat with a Wistar wild type (Wt) male (arrows). ( e ) Genotyping by PCR analysis of the extracted DNA from ear snips of the offspring. B: brown coat colour, W: white coat colour, V: CAG/human CD63-GFP vector, and M: size marker.

    Article Snippet: The plasmid CAG/human CD63-GFP was linearized by SalI and was then transfected into Wistar rESCs using a Mouse ES Cell Nucleofector Kit (Lonza, Basel, Swiss) and an Amaxa Nucleofector (A-013 program) as described previously . pCAG/human CD63-GFP contained a neomycin/kanamycin selection cassette.

    Techniques: Transgenic Assay, Construct, Transfection, Cell Culture, Derivative Assay, Injection, Polymerase Chain Reaction, Plasmid Preparation, Marker