sheep polyclonal anti human tgn46 Search Results


human tgn46  (Bio-Rad)
93
Bio-Rad human tgn46
Human Tgn46, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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polyclonal sheep anti tgn46 antibody  (Bio-Rad)
96
Bio-Rad polyclonal sheep anti tgn46 antibody
Polyclonal Sheep Anti Tgn46 Antibody, supplied by Bio-Rad, 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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igg1 sheep recombinant human tgn46 abdserotec if  (Bio-Rad)
90
Bio-Rad igg1 sheep recombinant human tgn46 abdserotec if
Igg1 Sheep Recombinant Human Tgn46 Abdserotec If, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sheep anti tgn46  (Bio-Rad)
93
Bio-Rad sheep anti tgn46
Sheep Anti Tgn46, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 1 article reviews
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1b5 mouse igg2b mark marsh tgn46 anti tgn46 sheep polyclonal abd serotec annexin  (Bio-Rad)
95
Bio-Rad 1b5 mouse igg2b mark marsh tgn46 anti tgn46 sheep polyclonal abd serotec annexin
Fig. 1. Subcellular localization of exocytic pathway markers Giantin and <t>TGN46</t> in HCMV-infected cells. BJ1 cells were either mock infected (upper panels) or infected with HCMV at an moi of 0.5 (lower panels). After 4 dpi, cells were fixed, permeabilized and stained with anti-Giantin (green in left panels), anti-TGN (green in right panels) and anti-HCMV glycoprotein gH (red) antibodies. In HCMV-infected cells Giantin surrounded the virus factory while TGN46 was accumulated within it. Scale bars, 20 mm.
1b5 Mouse Igg2b Mark Marsh Tgn46 Anti Tgn46 Sheep Polyclonal Abd Serotec Annexin, supplied by Bio-Rad, 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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sheep anti-human tgn46  (Serotech Inc)
90
Serotech Inc sheep anti-human tgn46
APP is cleaved by BACE1 independent of endosomal localization. (A) H4 human neuroglioma cells were transfected with constructs encoding APP-F/P-D/A or APP-F/P-D/A with additional mutation of all three tyrosine residues in the cytosolic tail (APP 3Y-F/P-D/A), both expressing C-terminal GFP. Cells were stained for <t>TGN46</t> and examined by fluorescence microscopy. Merging red and green channels generated the third picture on each row; yellow indicates overlapping localization. Insets show 3× magnifications. (B) Immunofluorescence images from A were quantified for Golgi localization. Values are the mean ± SD from three different experiments. ***p < 0.001. (C) Cells were biotinylated on the cell surface as described in and proteins analyzed by immunoblotting with an anti–GFP-HRP antibody. (D) Rat cortical neurons were cotransfected with APP 3Y-F/P-D/A and wild-type or mutant BACE1 constructs on DIV-4 and analyzed on DIV-7 as described in . Bar, 10 μm.
Sheep Anti Human Tgn46, supplied by Serotech Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti myc a14  (Bio-Rad)
92
Bio-Rad rabbit anti myc a14
APP is cleaved by BACE1 independent of endosomal localization. (A) H4 human neuroglioma cells were transfected with constructs encoding APP-F/P-D/A or APP-F/P-D/A with additional mutation of all three tyrosine residues in the cytosolic tail (APP 3Y-F/P-D/A), both expressing C-terminal GFP. Cells were stained for <t>TGN46</t> and examined by fluorescence microscopy. Merging red and green channels generated the third picture on each row; yellow indicates overlapping localization. Insets show 3× magnifications. (B) Immunofluorescence images from A were quantified for Golgi localization. Values are the mean ± SD from three different experiments. ***p < 0.001. (C) Cells were biotinylated on the cell surface as described in and proteins analyzed by immunoblotting with an anti–GFP-HRP antibody. (D) Rat cortical neurons were cotransfected with APP 3Y-F/P-D/A and wild-type or mutant BACE1 constructs on DIV-4 and analyzed on DIV-7 as described in . Bar, 10 μm.
Rabbit Anti Myc A14, supplied by Bio-Rad, 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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sheep anti-tgn46  (GeneTex)
90
GeneTex sheep anti-tgn46
APP is cleaved by BACE1 independent of endosomal localization. (A) H4 human neuroglioma cells were transfected with constructs encoding APP-F/P-D/A or APP-F/P-D/A with additional mutation of all three tyrosine residues in the cytosolic tail (APP 3Y-F/P-D/A), both expressing C-terminal GFP. Cells were stained for <t>TGN46</t> and examined by fluorescence microscopy. Merging red and green channels generated the third picture on each row; yellow indicates overlapping localization. Insets show 3× magnifications. (B) Immunofluorescence images from A were quantified for Golgi localization. Values are the mean ± SD from three different experiments. ***p < 0.001. (C) Cells were biotinylated on the cell surface as described in and proteins analyzed by immunoblotting with an anti–GFP-HRP antibody. (D) Rat cortical neurons were cotransfected with APP 3Y-F/P-D/A and wild-type or mutant BACE1 constructs on DIV-4 and analyzed on DIV-7 as described in . Bar, 10 μm.
Sheep Anti Tgn46, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse monoclonal antibody mab anti pp28  (Abcam)
99
Abcam mouse monoclonal antibody mab anti pp28
APP is cleaved by BACE1 independent of endosomal localization. (A) H4 human neuroglioma cells were transfected with constructs encoding APP-F/P-D/A or APP-F/P-D/A with additional mutation of all three tyrosine residues in the cytosolic tail (APP 3Y-F/P-D/A), both expressing C-terminal GFP. Cells were stained for <t>TGN46</t> and examined by fluorescence microscopy. Merging red and green channels generated the third picture on each row; yellow indicates overlapping localization. Insets show 3× magnifications. (B) Immunofluorescence images from A were quantified for Golgi localization. Values are the mean ± SD from three different experiments. ***p < 0.001. (C) Cells were biotinylated on the cell surface as described in and proteins analyzed by immunoblotting with an anti–GFP-HRP antibody. (D) Rat cortical neurons were cotransfected with APP 3Y-F/P-D/A and wild-type or mutant BACE1 constructs on DIV-4 and analyzed on DIV-7 as described in . Bar, 10 μm.
Mouse Monoclonal Antibody Mab Anti Pp28, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-ccr5 mab  (Becton Dickinson)
90
Becton Dickinson anti-ccr5 mab
APP is cleaved by BACE1 independent of endosomal localization. (A) H4 human neuroglioma cells were transfected with constructs encoding APP-F/P-D/A or APP-F/P-D/A with additional mutation of all three tyrosine residues in the cytosolic tail (APP 3Y-F/P-D/A), both expressing C-terminal GFP. Cells were stained for <t>TGN46</t> and examined by fluorescence microscopy. Merging red and green channels generated the third picture on each row; yellow indicates overlapping localization. Insets show 3× magnifications. (B) Immunofluorescence images from A were quantified for Golgi localization. Values are the mean ± SD from three different experiments. ***p < 0.001. (C) Cells were biotinylated on the cell surface as described in and proteins analyzed by immunoblotting with an anti–GFP-HRP antibody. (D) Rat cortical neurons were cotransfected with APP 3Y-F/P-D/A and wild-type or mutant BACE1 constructs on DIV-4 and analyzed on DIV-7 as described in . Bar, 10 μm.
Anti Ccr5 Mab, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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polyclonal antibodies (abs) for c-myc  (MBL Life science)
90
MBL Life science polyclonal antibodies (abs) for c-myc
APP is cleaved by BACE1 independent of endosomal localization. (A) H4 human neuroglioma cells were transfected with constructs encoding APP-F/P-D/A or APP-F/P-D/A with additional mutation of all three tyrosine residues in the cytosolic tail (APP 3Y-F/P-D/A), both expressing C-terminal GFP. Cells were stained for <t>TGN46</t> and examined by fluorescence microscopy. Merging red and green channels generated the third picture on each row; yellow indicates overlapping localization. Insets show 3× magnifications. (B) Immunofluorescence images from A were quantified for Golgi localization. Values are the mean ± SD from three different experiments. ***p < 0.001. (C) Cells were biotinylated on the cell surface as described in and proteins analyzed by immunoblotting with an anti–GFP-HRP antibody. (D) Rat cortical neurons were cotransfected with APP 3Y-F/P-D/A and wild-type or mutant BACE1 constructs on DIV-4 and analyzed on DIV-7 as described in . Bar, 10 μm.
Polyclonal Antibodies (Abs) For C Myc, supplied by MBL Life science, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Fig. 1. Subcellular localization of exocytic pathway markers Giantin and TGN46 in HCMV-infected cells. BJ1 cells were either mock infected (upper panels) or infected with HCMV at an moi of 0.5 (lower panels). After 4 dpi, cells were fixed, permeabilized and stained with anti-Giantin (green in left panels), anti-TGN (green in right panels) and anti-HCMV glycoprotein gH (red) antibodies. In HCMV-infected cells Giantin surrounded the virus factory while TGN46 was accumulated within it. Scale bars, 20 mm.

Journal: Cellular microbiology

Article Title: Human cytomegalovirus final envelopment on membranes containing both trans-Golgi network and endosomal markers.

doi: 10.1111/j.1462-5822.2009.01405.x

Figure Lengend Snippet: Fig. 1. Subcellular localization of exocytic pathway markers Giantin and TGN46 in HCMV-infected cells. BJ1 cells were either mock infected (upper panels) or infected with HCMV at an moi of 0.5 (lower panels). After 4 dpi, cells were fixed, permeabilized and stained with anti-Giantin (green in left panels), anti-TGN (green in right panels) and anti-HCMV glycoprotein gH (red) antibodies. In HCMV-infected cells Giantin surrounded the virus factory while TGN46 was accumulated within it. Scale bars, 20 mm.

Article Snippet: Antigen Antibodies name Species and isotype Source HCMV glycoprotein H HCMV 16 Mouse IgG1 Helena Browne HCMV glycoprotein B HCMV 37 Mouse IgG2a Helena Browne HCMV UL33 Anti-HCMV UL33 Rabbit polyclonal Wade Gibson HCMV pp28 Anti-HCMV pp28 Mouse IgG1 Thomas Shenk HSV-1 glycoprotein D AP7 Mouse IgG2a Helena Browne CD63 (lumenal domain) Anti-CD63, 1B5 Mouse IgG2b Mark Marsh TGN46 Anti-TGN46 Sheep polyclonal AbD-Serotec Annexin I Anti-annexin I Mouse IgG1 Claire Isacke Early endosome antigen 1 Anti-EEA1 Mouse IgG1 BD Biosciences Hepatocyte growth factor-regulated Tyr-kinase substrate Anti-HRS 958/3 Rabbit polyclonal Sylvie Urbé Transferrin receptor (lumenal domain) B3/25 Mouse IgG1 Santa Cruz Biotechnology Transferrin receptor (cytoplasmic tail) H68.4 Mouse IgG1 Zymed Laboratories CI-M6PR (lumenal domain) Anti-MPR300 G VII-3 Rabbit polyclonal Kurt von Figura CI-M6PR (cytoplasmic tail) Anti-MPR300 H-CI-II Rabbit polyclonal Kurt von Figura CD-M6PR (lumenal domain) Anti-MPR46 HP-GI-3 Rabbit polyclonal Kurt von Figura CD-M6PR (cytoplasmic tail) Anti-MPR46 MSCI Rabbit polyclonal Kurt von Figura CD-M6PR Anti-MPR46, 22d4 Mouse IgG2b DSHB b-actin Anti-b-actin AC-15 Mouse IgG1 Sigma Aldrich Actin Anti-actin Rabbit polyclonal Sigma Aldrich Clathrin heavy chain Anti-CHC 23 Mouse IgG1 BD Biosciences © 2009 Blackwell Publishing Ltd, Cellular Microbiology, 12, 386–404 0.5 infectious particles per cell.

Techniques: Infection, Staining, Virus

Fig. 5. Examination of cellular markers in cell lysates and supernatants of HCMV-infected cells. A. Equal number of BJ1 cells mock infected or HCMV infected for 4 days at an moi of 3 were lysed and viral particles pelleted by centrifugation of the clarified supernatants were mixed with non-reducing SDS-PAGE sample buffer. Lysates were separated by SDS-PAGE and transferred to PVDF membranes. Membranes were cut and incubated, or sequentially incubated, with antibodies against cellular markers and antibodies against clathrin heavy chain or actin to check protein loading in cell lysates, and antiviral tegument protein pp28 antibodies to check HCMV viral particles secreted into the supernatants. Autoradiography films were scanned; images were cropped and assembled with Adobe Photoshop. Molecular weights in kDa are indicated. We found CD63, TGN46, EEA1, TfR and CI-M6PR in the supernatants of HCMV-infected cells. B. Results of cell lysates in (A) were analysed by densitometry, normalized to the expression levels of loading controls and expressed as a percentage of the levels in uninfected cells. We observed a reproducible reduction in the levels of CD63, TGN46 and CD-M6PR together with an increase in the levels of TfR by comparison with uninfected cells.

Journal: Cellular microbiology

Article Title: Human cytomegalovirus final envelopment on membranes containing both trans-Golgi network and endosomal markers.

doi: 10.1111/j.1462-5822.2009.01405.x

Figure Lengend Snippet: Fig. 5. Examination of cellular markers in cell lysates and supernatants of HCMV-infected cells. A. Equal number of BJ1 cells mock infected or HCMV infected for 4 days at an moi of 3 were lysed and viral particles pelleted by centrifugation of the clarified supernatants were mixed with non-reducing SDS-PAGE sample buffer. Lysates were separated by SDS-PAGE and transferred to PVDF membranes. Membranes were cut and incubated, or sequentially incubated, with antibodies against cellular markers and antibodies against clathrin heavy chain or actin to check protein loading in cell lysates, and antiviral tegument protein pp28 antibodies to check HCMV viral particles secreted into the supernatants. Autoradiography films were scanned; images were cropped and assembled with Adobe Photoshop. Molecular weights in kDa are indicated. We found CD63, TGN46, EEA1, TfR and CI-M6PR in the supernatants of HCMV-infected cells. B. Results of cell lysates in (A) were analysed by densitometry, normalized to the expression levels of loading controls and expressed as a percentage of the levels in uninfected cells. We observed a reproducible reduction in the levels of CD63, TGN46 and CD-M6PR together with an increase in the levels of TfR by comparison with uninfected cells.

Article Snippet: Antigen Antibodies name Species and isotype Source HCMV glycoprotein H HCMV 16 Mouse IgG1 Helena Browne HCMV glycoprotein B HCMV 37 Mouse IgG2a Helena Browne HCMV UL33 Anti-HCMV UL33 Rabbit polyclonal Wade Gibson HCMV pp28 Anti-HCMV pp28 Mouse IgG1 Thomas Shenk HSV-1 glycoprotein D AP7 Mouse IgG2a Helena Browne CD63 (lumenal domain) Anti-CD63, 1B5 Mouse IgG2b Mark Marsh TGN46 Anti-TGN46 Sheep polyclonal AbD-Serotec Annexin I Anti-annexin I Mouse IgG1 Claire Isacke Early endosome antigen 1 Anti-EEA1 Mouse IgG1 BD Biosciences Hepatocyte growth factor-regulated Tyr-kinase substrate Anti-HRS 958/3 Rabbit polyclonal Sylvie Urbé Transferrin receptor (lumenal domain) B3/25 Mouse IgG1 Santa Cruz Biotechnology Transferrin receptor (cytoplasmic tail) H68.4 Mouse IgG1 Zymed Laboratories CI-M6PR (lumenal domain) Anti-MPR300 G VII-3 Rabbit polyclonal Kurt von Figura CI-M6PR (cytoplasmic tail) Anti-MPR300 H-CI-II Rabbit polyclonal Kurt von Figura CD-M6PR (lumenal domain) Anti-MPR46 HP-GI-3 Rabbit polyclonal Kurt von Figura CD-M6PR (cytoplasmic tail) Anti-MPR46 MSCI Rabbit polyclonal Kurt von Figura CD-M6PR Anti-MPR46, 22d4 Mouse IgG2b DSHB b-actin Anti-b-actin AC-15 Mouse IgG1 Sigma Aldrich Actin Anti-actin Rabbit polyclonal Sigma Aldrich Clathrin heavy chain Anti-CHC 23 Mouse IgG1 BD Biosciences © 2009 Blackwell Publishing Ltd, Cellular Microbiology, 12, 386–404 0.5 infectious particles per cell.

Techniques: Infection, Centrifugation, SDS Page, Incubation, Autoradiography, Expressing, Comparison

Fig. 6. Examination of cellular markers in purified HCMV virions. A. Electron microscopy of purified HCMV virions negatively stained with 2% uranyl acetate. When viral particles were partially disrupted, uranyl acetate revealed the nucleocapsids. Scale bar, 200 nm. B. 35 ng of purified virions per lane were separated by SDS-PAGE under non-reducing conditions and transferred to PVDF membranes. Membranes were cut and incubated, or sequentially incubated, with antibodies against cellular markers and antiviral tegument protein pp28 antibodies. Autoradiography films were scanned; images were cropped and assembled with Adobe Photoshop. Molecular weights in kDa are indicated. We found CD63, TGN46, TfR and CI-M6PR in purified virions.

Journal: Cellular microbiology

Article Title: Human cytomegalovirus final envelopment on membranes containing both trans-Golgi network and endosomal markers.

doi: 10.1111/j.1462-5822.2009.01405.x

Figure Lengend Snippet: Fig. 6. Examination of cellular markers in purified HCMV virions. A. Electron microscopy of purified HCMV virions negatively stained with 2% uranyl acetate. When viral particles were partially disrupted, uranyl acetate revealed the nucleocapsids. Scale bar, 200 nm. B. 35 ng of purified virions per lane were separated by SDS-PAGE under non-reducing conditions and transferred to PVDF membranes. Membranes were cut and incubated, or sequentially incubated, with antibodies against cellular markers and antiviral tegument protein pp28 antibodies. Autoradiography films were scanned; images were cropped and assembled with Adobe Photoshop. Molecular weights in kDa are indicated. We found CD63, TGN46, TfR and CI-M6PR in purified virions.

Article Snippet: Antigen Antibodies name Species and isotype Source HCMV glycoprotein H HCMV 16 Mouse IgG1 Helena Browne HCMV glycoprotein B HCMV 37 Mouse IgG2a Helena Browne HCMV UL33 Anti-HCMV UL33 Rabbit polyclonal Wade Gibson HCMV pp28 Anti-HCMV pp28 Mouse IgG1 Thomas Shenk HSV-1 glycoprotein D AP7 Mouse IgG2a Helena Browne CD63 (lumenal domain) Anti-CD63, 1B5 Mouse IgG2b Mark Marsh TGN46 Anti-TGN46 Sheep polyclonal AbD-Serotec Annexin I Anti-annexin I Mouse IgG1 Claire Isacke Early endosome antigen 1 Anti-EEA1 Mouse IgG1 BD Biosciences Hepatocyte growth factor-regulated Tyr-kinase substrate Anti-HRS 958/3 Rabbit polyclonal Sylvie Urbé Transferrin receptor (lumenal domain) B3/25 Mouse IgG1 Santa Cruz Biotechnology Transferrin receptor (cytoplasmic tail) H68.4 Mouse IgG1 Zymed Laboratories CI-M6PR (lumenal domain) Anti-MPR300 G VII-3 Rabbit polyclonal Kurt von Figura CI-M6PR (cytoplasmic tail) Anti-MPR300 H-CI-II Rabbit polyclonal Kurt von Figura CD-M6PR (lumenal domain) Anti-MPR46 HP-GI-3 Rabbit polyclonal Kurt von Figura CD-M6PR (cytoplasmic tail) Anti-MPR46 MSCI Rabbit polyclonal Kurt von Figura CD-M6PR Anti-MPR46, 22d4 Mouse IgG2b DSHB b-actin Anti-b-actin AC-15 Mouse IgG1 Sigma Aldrich Actin Anti-actin Rabbit polyclonal Sigma Aldrich Clathrin heavy chain Anti-CHC 23 Mouse IgG1 BD Biosciences © 2009 Blackwell Publishing Ltd, Cellular Microbiology, 12, 386–404 0.5 infectious particles per cell.

Techniques: Electron Microscopy, Staining, SDS Page, Incubation, Autoradiography

Fig. 7. Transcriptional regulation of cellular markers during HCMV infection. Equal number of BJ1 cells were mock infected or infected with HCMV at an moi of 3, and RNA was extracted at the indicated times. Gene expression of cellular markers and HCMV UL83 (viral protein pp65) was measured by qPCR, normalized to 18S rRNA expression and calibrated to the levels in uninfected cells. CD63, TGN46, HRS and annexin I transcripts were downregulated over the time-course of HCMV infection, while EEA1 and CI-M6PR transcripts levels changed slightly. TfR and CD-M6PR transcripts were transiently upregulated at 1 dpi and then downregulated with the mRNA levels of TfR increasing at 5 dpi and of CD-M6PR unaltered after 3 dpi. HCMV pp65 transcripts levels increased during the infection. Data corresponding to three biological replicates were averaged, bars represent the average normalized fold increase, and error bars represent standard errors of the means.

Journal: Cellular microbiology

Article Title: Human cytomegalovirus final envelopment on membranes containing both trans-Golgi network and endosomal markers.

doi: 10.1111/j.1462-5822.2009.01405.x

Figure Lengend Snippet: Fig. 7. Transcriptional regulation of cellular markers during HCMV infection. Equal number of BJ1 cells were mock infected or infected with HCMV at an moi of 3, and RNA was extracted at the indicated times. Gene expression of cellular markers and HCMV UL83 (viral protein pp65) was measured by qPCR, normalized to 18S rRNA expression and calibrated to the levels in uninfected cells. CD63, TGN46, HRS and annexin I transcripts were downregulated over the time-course of HCMV infection, while EEA1 and CI-M6PR transcripts levels changed slightly. TfR and CD-M6PR transcripts were transiently upregulated at 1 dpi and then downregulated with the mRNA levels of TfR increasing at 5 dpi and of CD-M6PR unaltered after 3 dpi. HCMV pp65 transcripts levels increased during the infection. Data corresponding to three biological replicates were averaged, bars represent the average normalized fold increase, and error bars represent standard errors of the means.

Article Snippet: Antigen Antibodies name Species and isotype Source HCMV glycoprotein H HCMV 16 Mouse IgG1 Helena Browne HCMV glycoprotein B HCMV 37 Mouse IgG2a Helena Browne HCMV UL33 Anti-HCMV UL33 Rabbit polyclonal Wade Gibson HCMV pp28 Anti-HCMV pp28 Mouse IgG1 Thomas Shenk HSV-1 glycoprotein D AP7 Mouse IgG2a Helena Browne CD63 (lumenal domain) Anti-CD63, 1B5 Mouse IgG2b Mark Marsh TGN46 Anti-TGN46 Sheep polyclonal AbD-Serotec Annexin I Anti-annexin I Mouse IgG1 Claire Isacke Early endosome antigen 1 Anti-EEA1 Mouse IgG1 BD Biosciences Hepatocyte growth factor-regulated Tyr-kinase substrate Anti-HRS 958/3 Rabbit polyclonal Sylvie Urbé Transferrin receptor (lumenal domain) B3/25 Mouse IgG1 Santa Cruz Biotechnology Transferrin receptor (cytoplasmic tail) H68.4 Mouse IgG1 Zymed Laboratories CI-M6PR (lumenal domain) Anti-MPR300 G VII-3 Rabbit polyclonal Kurt von Figura CI-M6PR (cytoplasmic tail) Anti-MPR300 H-CI-II Rabbit polyclonal Kurt von Figura CD-M6PR (lumenal domain) Anti-MPR46 HP-GI-3 Rabbit polyclonal Kurt von Figura CD-M6PR (cytoplasmic tail) Anti-MPR46 MSCI Rabbit polyclonal Kurt von Figura CD-M6PR Anti-MPR46, 22d4 Mouse IgG2b DSHB b-actin Anti-b-actin AC-15 Mouse IgG1 Sigma Aldrich Actin Anti-actin Rabbit polyclonal Sigma Aldrich Clathrin heavy chain Anti-CHC 23 Mouse IgG1 BD Biosciences © 2009 Blackwell Publishing Ltd, Cellular Microbiology, 12, 386–404 0.5 infectious particles per cell.

Techniques: Infection, Gene Expression, Expressing

Fig. 8. Immuno-gold localization of viral and cellular proteins on the viral envelope of isolated HCMV viral particles. Isolated viral particles were labelled with antibodies against HCMV viral glycoprotein gH (A, PAG10) and viral chemokine receptor-like protein UL33 (E, PAG15), TGN46 (B, PAG15), CD63 (C, PAG10), EEA1 (F, PAG10), annexin1 (G, PAG10), TfR lumenal domain (H, PAG10), CI-M6PR lumenal domain (I, PAG10) either without (A–D, H–I) or after permeabilization with saponin (E–G). (D) Double-labelling with antibodies against CD63 (PAG10) and TGN46 (PAG15). When viral particles were partially disrupted, uranyl acetate revealed the nucleocapsids (A, C, E–G). Scale bar, 50 nm.

Journal: Cellular microbiology

Article Title: Human cytomegalovirus final envelopment on membranes containing both trans-Golgi network and endosomal markers.

doi: 10.1111/j.1462-5822.2009.01405.x

Figure Lengend Snippet: Fig. 8. Immuno-gold localization of viral and cellular proteins on the viral envelope of isolated HCMV viral particles. Isolated viral particles were labelled with antibodies against HCMV viral glycoprotein gH (A, PAG10) and viral chemokine receptor-like protein UL33 (E, PAG15), TGN46 (B, PAG15), CD63 (C, PAG10), EEA1 (F, PAG10), annexin1 (G, PAG10), TfR lumenal domain (H, PAG10), CI-M6PR lumenal domain (I, PAG10) either without (A–D, H–I) or after permeabilization with saponin (E–G). (D) Double-labelling with antibodies against CD63 (PAG10) and TGN46 (PAG15). When viral particles were partially disrupted, uranyl acetate revealed the nucleocapsids (A, C, E–G). Scale bar, 50 nm.

Article Snippet: Antigen Antibodies name Species and isotype Source HCMV glycoprotein H HCMV 16 Mouse IgG1 Helena Browne HCMV glycoprotein B HCMV 37 Mouse IgG2a Helena Browne HCMV UL33 Anti-HCMV UL33 Rabbit polyclonal Wade Gibson HCMV pp28 Anti-HCMV pp28 Mouse IgG1 Thomas Shenk HSV-1 glycoprotein D AP7 Mouse IgG2a Helena Browne CD63 (lumenal domain) Anti-CD63, 1B5 Mouse IgG2b Mark Marsh TGN46 Anti-TGN46 Sheep polyclonal AbD-Serotec Annexin I Anti-annexin I Mouse IgG1 Claire Isacke Early endosome antigen 1 Anti-EEA1 Mouse IgG1 BD Biosciences Hepatocyte growth factor-regulated Tyr-kinase substrate Anti-HRS 958/3 Rabbit polyclonal Sylvie Urbé Transferrin receptor (lumenal domain) B3/25 Mouse IgG1 Santa Cruz Biotechnology Transferrin receptor (cytoplasmic tail) H68.4 Mouse IgG1 Zymed Laboratories CI-M6PR (lumenal domain) Anti-MPR300 G VII-3 Rabbit polyclonal Kurt von Figura CI-M6PR (cytoplasmic tail) Anti-MPR300 H-CI-II Rabbit polyclonal Kurt von Figura CD-M6PR (lumenal domain) Anti-MPR46 HP-GI-3 Rabbit polyclonal Kurt von Figura CD-M6PR (cytoplasmic tail) Anti-MPR46 MSCI Rabbit polyclonal Kurt von Figura CD-M6PR Anti-MPR46, 22d4 Mouse IgG2b DSHB b-actin Anti-b-actin AC-15 Mouse IgG1 Sigma Aldrich Actin Anti-actin Rabbit polyclonal Sigma Aldrich Clathrin heavy chain Anti-CHC 23 Mouse IgG1 BD Biosciences © 2009 Blackwell Publishing Ltd, Cellular Microbiology, 12, 386–404 0.5 infectious particles per cell.

Techniques: Isolation

Fig. 9. Immunoprecipitation of virus particles with anti-TGN46 and anti-CD63 antibodies. Viruses secreted into the cell medium were incubated at 4°C overnight with antibodies against HSV-1 gD (negative control), HCMV gH (positive control), TGN46 and CD63. The virus-antibodies mixture was incubated with Pansorbin cells and viruses were precipitated. Supernatans were added to fresh BJ1 cells to determine the number of infectious unprecipitated viruses as described in Experimental procedures. To calculate the percentage of infectivity, the infectivity of samples treated with anti-HSV-1 gD antibodies was set at 100%. Data corresponding to three experiments were averaged, bars represent the average relative units and error bars represent standard errors of the means.

Journal: Cellular microbiology

Article Title: Human cytomegalovirus final envelopment on membranes containing both trans-Golgi network and endosomal markers.

doi: 10.1111/j.1462-5822.2009.01405.x

Figure Lengend Snippet: Fig. 9. Immunoprecipitation of virus particles with anti-TGN46 and anti-CD63 antibodies. Viruses secreted into the cell medium were incubated at 4°C overnight with antibodies against HSV-1 gD (negative control), HCMV gH (positive control), TGN46 and CD63. The virus-antibodies mixture was incubated with Pansorbin cells and viruses were precipitated. Supernatans were added to fresh BJ1 cells to determine the number of infectious unprecipitated viruses as described in Experimental procedures. To calculate the percentage of infectivity, the infectivity of samples treated with anti-HSV-1 gD antibodies was set at 100%. Data corresponding to three experiments were averaged, bars represent the average relative units and error bars represent standard errors of the means.

Article Snippet: Antigen Antibodies name Species and isotype Source HCMV glycoprotein H HCMV 16 Mouse IgG1 Helena Browne HCMV glycoprotein B HCMV 37 Mouse IgG2a Helena Browne HCMV UL33 Anti-HCMV UL33 Rabbit polyclonal Wade Gibson HCMV pp28 Anti-HCMV pp28 Mouse IgG1 Thomas Shenk HSV-1 glycoprotein D AP7 Mouse IgG2a Helena Browne CD63 (lumenal domain) Anti-CD63, 1B5 Mouse IgG2b Mark Marsh TGN46 Anti-TGN46 Sheep polyclonal AbD-Serotec Annexin I Anti-annexin I Mouse IgG1 Claire Isacke Early endosome antigen 1 Anti-EEA1 Mouse IgG1 BD Biosciences Hepatocyte growth factor-regulated Tyr-kinase substrate Anti-HRS 958/3 Rabbit polyclonal Sylvie Urbé Transferrin receptor (lumenal domain) B3/25 Mouse IgG1 Santa Cruz Biotechnology Transferrin receptor (cytoplasmic tail) H68.4 Mouse IgG1 Zymed Laboratories CI-M6PR (lumenal domain) Anti-MPR300 G VII-3 Rabbit polyclonal Kurt von Figura CI-M6PR (cytoplasmic tail) Anti-MPR300 H-CI-II Rabbit polyclonal Kurt von Figura CD-M6PR (lumenal domain) Anti-MPR46 HP-GI-3 Rabbit polyclonal Kurt von Figura CD-M6PR (cytoplasmic tail) Anti-MPR46 MSCI Rabbit polyclonal Kurt von Figura CD-M6PR Anti-MPR46, 22d4 Mouse IgG2b DSHB b-actin Anti-b-actin AC-15 Mouse IgG1 Sigma Aldrich Actin Anti-actin Rabbit polyclonal Sigma Aldrich Clathrin heavy chain Anti-CHC 23 Mouse IgG1 BD Biosciences © 2009 Blackwell Publishing Ltd, Cellular Microbiology, 12, 386–404 0.5 infectious particles per cell.

Techniques: Immunoprecipitation, Virus, Incubation, Negative Control, Positive Control, Infection

Fig. 10. Model of HCMV envelopment and secretion. Nucleocapsids assembled in the nucleus are released into the cytoplasm through the nuclear membranes. In the perinuclear region, where virus factories are established, nucleocapsids acquire tegument before completing their final envelopment in a hybrid compartment or in the transport vesicles between the TGN and endosomes. During envelopment viral glycoproteins and TGN46, CD63, annexin 1, EEA1, TfR and CI-M6PR are incorporated into the viral membrane of numerous particles. Mature virions are transported inside a vacuole towards the plasma membrane to be secreted into the extracellular environment. N: nucleus, INM: inner nuclear membrane, ONM: outer nuclear membrane, ER, endoplasmic reticulum; TGN, trans-Golgi network; MVB, multivesicular body; EE, early endosome; V, vacuole; PM, plasma membrane.

Journal: Cellular microbiology

Article Title: Human cytomegalovirus final envelopment on membranes containing both trans-Golgi network and endosomal markers.

doi: 10.1111/j.1462-5822.2009.01405.x

Figure Lengend Snippet: Fig. 10. Model of HCMV envelopment and secretion. Nucleocapsids assembled in the nucleus are released into the cytoplasm through the nuclear membranes. In the perinuclear region, where virus factories are established, nucleocapsids acquire tegument before completing their final envelopment in a hybrid compartment or in the transport vesicles between the TGN and endosomes. During envelopment viral glycoproteins and TGN46, CD63, annexin 1, EEA1, TfR and CI-M6PR are incorporated into the viral membrane of numerous particles. Mature virions are transported inside a vacuole towards the plasma membrane to be secreted into the extracellular environment. N: nucleus, INM: inner nuclear membrane, ONM: outer nuclear membrane, ER, endoplasmic reticulum; TGN, trans-Golgi network; MVB, multivesicular body; EE, early endosome; V, vacuole; PM, plasma membrane.

Article Snippet: Antigen Antibodies name Species and isotype Source HCMV glycoprotein H HCMV 16 Mouse IgG1 Helena Browne HCMV glycoprotein B HCMV 37 Mouse IgG2a Helena Browne HCMV UL33 Anti-HCMV UL33 Rabbit polyclonal Wade Gibson HCMV pp28 Anti-HCMV pp28 Mouse IgG1 Thomas Shenk HSV-1 glycoprotein D AP7 Mouse IgG2a Helena Browne CD63 (lumenal domain) Anti-CD63, 1B5 Mouse IgG2b Mark Marsh TGN46 Anti-TGN46 Sheep polyclonal AbD-Serotec Annexin I Anti-annexin I Mouse IgG1 Claire Isacke Early endosome antigen 1 Anti-EEA1 Mouse IgG1 BD Biosciences Hepatocyte growth factor-regulated Tyr-kinase substrate Anti-HRS 958/3 Rabbit polyclonal Sylvie Urbé Transferrin receptor (lumenal domain) B3/25 Mouse IgG1 Santa Cruz Biotechnology Transferrin receptor (cytoplasmic tail) H68.4 Mouse IgG1 Zymed Laboratories CI-M6PR (lumenal domain) Anti-MPR300 G VII-3 Rabbit polyclonal Kurt von Figura CI-M6PR (cytoplasmic tail) Anti-MPR300 H-CI-II Rabbit polyclonal Kurt von Figura CD-M6PR (lumenal domain) Anti-MPR46 HP-GI-3 Rabbit polyclonal Kurt von Figura CD-M6PR (cytoplasmic tail) Anti-MPR46 MSCI Rabbit polyclonal Kurt von Figura CD-M6PR Anti-MPR46, 22d4 Mouse IgG2b DSHB b-actin Anti-b-actin AC-15 Mouse IgG1 Sigma Aldrich Actin Anti-actin Rabbit polyclonal Sigma Aldrich Clathrin heavy chain Anti-CHC 23 Mouse IgG1 BD Biosciences © 2009 Blackwell Publishing Ltd, Cellular Microbiology, 12, 386–404 0.5 infectious particles per cell.

Techniques: Virus, Membrane, Clinical Proteomics

APP is cleaved by BACE1 independent of endosomal localization. (A) H4 human neuroglioma cells were transfected with constructs encoding APP-F/P-D/A or APP-F/P-D/A with additional mutation of all three tyrosine residues in the cytosolic tail (APP 3Y-F/P-D/A), both expressing C-terminal GFP. Cells were stained for TGN46 and examined by fluorescence microscopy. Merging red and green channels generated the third picture on each row; yellow indicates overlapping localization. Insets show 3× magnifications. (B) Immunofluorescence images from A were quantified for Golgi localization. Values are the mean ± SD from three different experiments. ***p < 0.001. (C) Cells were biotinylated on the cell surface as described in and proteins analyzed by immunoblotting with an anti–GFP-HRP antibody. (D) Rat cortical neurons were cotransfected with APP 3Y-F/P-D/A and wild-type or mutant BACE1 constructs on DIV-4 and analyzed on DIV-7 as described in . Bar, 10 μm.

Journal: Molecular Biology of the Cell

Article Title: Adaptor protein 2–mediated endocytosis of the β-secretase BACE1 is dispensable for amyloid precursor protein processing

doi: 10.1091/mbc.E11-11-0944

Figure Lengend Snippet: APP is cleaved by BACE1 independent of endosomal localization. (A) H4 human neuroglioma cells were transfected with constructs encoding APP-F/P-D/A or APP-F/P-D/A with additional mutation of all three tyrosine residues in the cytosolic tail (APP 3Y-F/P-D/A), both expressing C-terminal GFP. Cells were stained for TGN46 and examined by fluorescence microscopy. Merging red and green channels generated the third picture on each row; yellow indicates overlapping localization. Insets show 3× magnifications. (B) Immunofluorescence images from A were quantified for Golgi localization. Values are the mean ± SD from three different experiments. ***p < 0.001. (C) Cells were biotinylated on the cell surface as described in and proteins analyzed by immunoblotting with an anti–GFP-HRP antibody. (D) Rat cortical neurons were cotransfected with APP 3Y-F/P-D/A and wild-type or mutant BACE1 constructs on DIV-4 and analyzed on DIV-7 as described in . Bar, 10 μm.

Article Snippet: Sheep anti-human TGN46 from Serotech (Raleigh, NC) was used.

Techniques: Transfection, Construct, Mutagenesis, Expressing, Staining, Fluorescence, Microscopy, Generated, Immunofluorescence, Western Blot