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retroviral packaging cell line  (ATCC)
92
ATCC retroviral packaging cell line
Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Retroviral Packaging Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sorvall x4r pro centrifuge  (Thermo Fisher)
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Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Sorvall X4r Pro Centrifuge, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cell culture plates  (Thermo Fisher)
96
Thermo Fisher cell culture plates
Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Cell Culture Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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zinc chloride  (Chem Impex International)
95
Chem Impex International zinc chloride
Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Zinc Chloride, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cell suspension to centrifuge  (Thermo Fisher)
94
Thermo Fisher cell suspension to centrifuge
Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Cell Suspension To Centrifuge, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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centrifuge  (Thermo Fisher)
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Thermo Fisher centrifuge
Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Centrifuge, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sl 1 cells  (Thermo Fisher)
91
Thermo Fisher sl 1 cells
Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Sl 1 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cell culture ingredients  (Thermo Fisher)
94
Thermo Fisher cell culture ingredients
Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Cell Culture Ingredients, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sorvall st plus series centrifuge  (Thermo Fisher)
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Thermo Fisher sorvall st plus series centrifuge
Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Sorvall St Plus Series Centrifuge, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sorvall legent rt plus centrifuge  (Thermo Fisher)
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Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
Sorvall Legent Rt Plus Centrifuge, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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thermo scientific st8r  (Thermo Fisher)
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Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
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multifuge x pro series benchtop centrifuge housing  (Thermo Fisher)
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Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a <t>retroviral</t> vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.
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Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a retroviral vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.

Journal: Virology

Article Title: Characterization of the Epstein-Barr virus glycoprotein BMRF-2.

doi: 10.1016/j.virol.2006.09.047

Figure Lengend Snippet: Fig. 8. Glycosylation of BMRF-2. (A) HSC3sort cells were transduced with a retroviral vector expressing BMRF-2-GFP. After 48 h, cells were fixed and co- immunostained for BMRF-2 with ER or Golgi markers. ER and Golgi were detected with staining for GRP94 and Rhodamine-Lens Culinaris Agglutinin, respectively. BMRF-2 is shown in green fluorescence as a GFP fusion protein and the ER and Golgi markers are shown in red fluorescence. Yellow indicates colocalization of BMRF-2 with ER or Golgi markers. (B) The 293T cells were transfected with the BMRF-2 gene for 48 h and the membrane fractions of these cells were isolated. In a parallel experiment, total proteins from purified EBV virions were extracted. Transfected cells and virion samples were treated with a combination of neuraminidase and O-glycosidase, and with O-glycosidase and neuraminidase independently. Untreated samples served as a control. Treated and untreated samples were subjected to Western blot analysis using rat anti- BMRF-2 immune serum and proteins were separated in 7 M urea gels. Neu, neuraminidase; OG, O-glycosydase. Cells, 293T cells transfected with BMRFQ2. Virions, gradient-purified EBV virions.

Article Snippet: A human embryonic kidney cell line (293T) and a retroviral packaging cell line (ProPak A.52) were purchased from American Type Culture Collection (ATCC,Manassas, Virginia).

Techniques: Glycoproteomics, Transduction, Retroviral, Plasmid Preparation, Expressing, Staining, Fluorescence, Transfection, Membrane, Isolation, Purification, Control, Western Blot