cat 66002 1 ig Search Results


anti gfp  (Proteintech)
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gfp  (Proteintech)
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Proteintech gfp
Gfp, supplied by Proteintech, 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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cat 66002 1 ig  (Proteintech)
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Proteintech cat 66002 1 ig
Cat 66002 1 Ig, supplied by Proteintech, 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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mouse monoclonal anti gfp  (Proteintech)
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Mouse Monoclonal Anti Gfp, supplied by Proteintech, 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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antibody anti egfp mouse monoclonal santa cruz  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology antibody anti egfp mouse monoclonal santa cruz
Figure 2. W383 and FDRxLYL motifs act as CLSs on the catalytic domain surface. (a) Cilia localization of the indicated FDRxLYL mutants of <t>EGFP-INPP5E</t> was analyzed in hTERT-RPE1 cells as in Figure 1. Scale bars, 5 µm. (b) Magnification from INPP5E structure (PDB ID: 2xsw) showing the FDRxLYL motif residues (pink) and adjacent catalytic domain residues shown here to affect ciliary targeting (green). Distance between W383 and F609 is indicated in angstroms. Beta-sheets and alpha-helices are shown as yellow and orange ribbons, respectively. Notice active site region on top left (cyan). (c) Cilia localization of the indicated EGFP-INPP5E constructs was analyzed as in (a). In both cases, images are representative of at least two independent experiments per construct, with >30 transfected-cell cilia visualized per construct and experiment. Scale bars, 5 µm. (d) Percentage of positive cilia
Antibody Anti Egfp Mouse Monoclonal Santa Cruz, supplied by Santa Cruz Biotechnology, 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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Image Search Results


Figure 2. W383 and FDRxLYL motifs act as CLSs on the catalytic domain surface. (a) Cilia localization of the indicated FDRxLYL mutants of EGFP-INPP5E was analyzed in hTERT-RPE1 cells as in Figure 1. Scale bars, 5 µm. (b) Magnification from INPP5E structure (PDB ID: 2xsw) showing the FDRxLYL motif residues (pink) and adjacent catalytic domain residues shown here to affect ciliary targeting (green). Distance between W383 and F609 is indicated in angstroms. Beta-sheets and alpha-helices are shown as yellow and orange ribbons, respectively. Notice active site region on top left (cyan). (c) Cilia localization of the indicated EGFP-INPP5E constructs was analyzed as in (a). In both cases, images are representative of at least two independent experiments per construct, with >30 transfected-cell cilia visualized per construct and experiment. Scale bars, 5 µm. (d) Percentage of positive cilia

Journal: eLife

Article Title: Multiple ciliary localization signals control INPP5E ciliary targeting

doi: 10.7554/elife.78383

Figure Lengend Snippet: Figure 2. W383 and FDRxLYL motifs act as CLSs on the catalytic domain surface. (a) Cilia localization of the indicated FDRxLYL mutants of EGFP-INPP5E was analyzed in hTERT-RPE1 cells as in Figure 1. Scale bars, 5 µm. (b) Magnification from INPP5E structure (PDB ID: 2xsw) showing the FDRxLYL motif residues (pink) and adjacent catalytic domain residues shown here to affect ciliary targeting (green). Distance between W383 and F609 is indicated in angstroms. Beta-sheets and alpha-helices are shown as yellow and orange ribbons, respectively. Notice active site region on top left (cyan). (c) Cilia localization of the indicated EGFP-INPP5E constructs was analyzed as in (a). In both cases, images are representative of at least two independent experiments per construct, with >30 transfected-cell cilia visualized per construct and experiment. Scale bars, 5 µm. (d) Percentage of positive cilia

Article Snippet: DOI: https://doi.org/10.7554/eLife.78383 27 of 43 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Cell line (Homo sapiens) Puromycin- sensitive hTERT- RPE1 This study Puromycin Acetyltransferase (PAC)KO cells Also used in Gonçalves et al., 2021 Cell line (Homo sapiens) INPP5E- KO hTERTRPE1 This study Clone 3 Derived from puromycin- sensitive hTERT- RPE1 Cell line (Homo sapiens) INPP5E- KO hTERTRPE1 This study Clone 12 Derived from puromycin- sensitive hTERT- RPE1 Cell line (Homo sapiens) 293T ATCC Cat # CRL- 3216 Derived from human embryonic kidney Cell line (Homo sapiens) HeLa ATCC Cat # CCL- 2 Derived from cervical carcinoma Cell line (Homo sapiens) Jurkat, Clone E6- 1 ATCC Cat # TIB- 152 T lymphoblasts from acute T cell leukemia Cell line (Homo sapiens) Raji ATCC Cat # CCL- 86 Lymphoblast- like cells from Burkitt’s lymphoma Cell line (Mus musculus) Csnk2a1- WT MEFs Loukil et al., 2021 N/A Control mouse embryonic fibroblasts (MEFs) Cell line (Mus musculus) Csnk2a1- KO MEFs Loukil et al., 2021 N/A Casein kinase 2 subunit alpha- null MEFs Antibody Anti- acetylated α-tubulin (mouse monoclonal) Sigma- Aldrich (Merck) Cat # T7451 (clone 6- 11B- 1) IF: 1:10,000 Antibody Anti-α-tubulin (mouse monoclonal) Proteintech Cat # 66031–1- Ig WB: 1:1000 Antibody Anti-γ-tubulin (mouse monoclonal) Santa Cruz Cat # sc- 17787 IF: 1:200 Antibody Anti- EGFP (mouse monoclonal) Proteintech Cat # 66002–1- Ig WB: 1:1000 Antibody Anti- EGFP (mouse monoclonal) Santa Cruz Cat # sc- 9996 IF: 1:200 Antibody Anti- ARL13B (mouse monoclonal) Proteintech Cat # 66739–1- Ig WB: 1:1000 Antibody Anti- Flag (mouse monoclonal) Sigma- Aldrich (Merck) Cat # F3165 (clone M2) WB: 1:2000 Antibody Anti- polyglutamylated tubulin (mouse monoclonal) Adipogen Cat # GT335 IF: 1:2000 Antibody Anti- EGFP (rabbit polyclonal) Proteintech Cat # 50430–2- AP IF: 1:200 WB: 1:1000 Antibody Anti- RPGR (rabbit polyclonal) Proteintech Cat # 16891–1- AP WB: 1:1000 Antibody Anti- TULP3 (rabbit polyclonal) Proteintech Cat # 13637–1- AP IF: 1:750 WB: 1:2000 Antibody Anti- CEP164 (rabbit polyclonal) Proteintech Cat # 22227–1- AP WB: 1:1000 Antibody Anti- ATG16L (rabbit polyclonal) MBL Cat # PM040 WB: 1:1000 Continued on next page Continued Cilleros- Rodriguez, Martin- Morales et al. eLife 2022;11:e78383.

Techniques: Construct, Transfection

Figure 6. CLS4 promotes INPP5E binding to PDE6D, RPGR and ARL13B. (a) The indicated EGFP-INPP5E variants were coexpressed in HEK293T cells with Flag-PDE6D, as indicated. Lysates were immunoprecipitated with GFP-Trap beads and analyzed by Western blot with the indicated antibodies. Molecular weight markers in kilodaltons are shown on the right. Quantitation of Flag-PDE6D co-immunoprecipitation (co-IP), as percentage relative to WT, is shown at the bottom. Quantitations are normalized relative to both immunoprecipitated EGFP constructs and lysate amounts of Flag-PDE6D. (b)

Journal: eLife

Article Title: Multiple ciliary localization signals control INPP5E ciliary targeting

doi: 10.7554/elife.78383

Figure Lengend Snippet: Figure 6. CLS4 promotes INPP5E binding to PDE6D, RPGR and ARL13B. (a) The indicated EGFP-INPP5E variants were coexpressed in HEK293T cells with Flag-PDE6D, as indicated. Lysates were immunoprecipitated with GFP-Trap beads and analyzed by Western blot with the indicated antibodies. Molecular weight markers in kilodaltons are shown on the right. Quantitation of Flag-PDE6D co-immunoprecipitation (co-IP), as percentage relative to WT, is shown at the bottom. Quantitations are normalized relative to both immunoprecipitated EGFP constructs and lysate amounts of Flag-PDE6D. (b)

Article Snippet: DOI: https://doi.org/10.7554/eLife.78383 27 of 43 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Cell line (Homo sapiens) Puromycin- sensitive hTERT- RPE1 This study Puromycin Acetyltransferase (PAC)KO cells Also used in Gonçalves et al., 2021 Cell line (Homo sapiens) INPP5E- KO hTERTRPE1 This study Clone 3 Derived from puromycin- sensitive hTERT- RPE1 Cell line (Homo sapiens) INPP5E- KO hTERTRPE1 This study Clone 12 Derived from puromycin- sensitive hTERT- RPE1 Cell line (Homo sapiens) 293T ATCC Cat # CRL- 3216 Derived from human embryonic kidney Cell line (Homo sapiens) HeLa ATCC Cat # CCL- 2 Derived from cervical carcinoma Cell line (Homo sapiens) Jurkat, Clone E6- 1 ATCC Cat # TIB- 152 T lymphoblasts from acute T cell leukemia Cell line (Homo sapiens) Raji ATCC Cat # CCL- 86 Lymphoblast- like cells from Burkitt’s lymphoma Cell line (Mus musculus) Csnk2a1- WT MEFs Loukil et al., 2021 N/A Control mouse embryonic fibroblasts (MEFs) Cell line (Mus musculus) Csnk2a1- KO MEFs Loukil et al., 2021 N/A Casein kinase 2 subunit alpha- null MEFs Antibody Anti- acetylated α-tubulin (mouse monoclonal) Sigma- Aldrich (Merck) Cat # T7451 (clone 6- 11B- 1) IF: 1:10,000 Antibody Anti-α-tubulin (mouse monoclonal) Proteintech Cat # 66031–1- Ig WB: 1:1000 Antibody Anti-γ-tubulin (mouse monoclonal) Santa Cruz Cat # sc- 17787 IF: 1:200 Antibody Anti- EGFP (mouse monoclonal) Proteintech Cat # 66002–1- Ig WB: 1:1000 Antibody Anti- EGFP (mouse monoclonal) Santa Cruz Cat # sc- 9996 IF: 1:200 Antibody Anti- ARL13B (mouse monoclonal) Proteintech Cat # 66739–1- Ig WB: 1:1000 Antibody Anti- Flag (mouse monoclonal) Sigma- Aldrich (Merck) Cat # F3165 (clone M2) WB: 1:2000 Antibody Anti- polyglutamylated tubulin (mouse monoclonal) Adipogen Cat # GT335 IF: 1:2000 Antibody Anti- EGFP (rabbit polyclonal) Proteintech Cat # 50430–2- AP IF: 1:200 WB: 1:1000 Antibody Anti- RPGR (rabbit polyclonal) Proteintech Cat # 16891–1- AP WB: 1:1000 Antibody Anti- TULP3 (rabbit polyclonal) Proteintech Cat # 13637–1- AP IF: 1:750 WB: 1:2000 Antibody Anti- CEP164 (rabbit polyclonal) Proteintech Cat # 22227–1- AP WB: 1:1000 Antibody Anti- ATG16L (rabbit polyclonal) MBL Cat # PM040 WB: 1:1000 Continued on next page Continued Cilleros- Rodriguez, Martin- Morales et al. eLife 2022;11:e78383.

Techniques: Binding Assay, Immunoprecipitation, Western Blot, Molecular Weight, Quantitation Assay, Co-Immunoprecipitation Assay, Construct

Figure 7. CLS2 and CLS3 regulate INPP5E binding to TULP3 and CEP164. (a) The indicated EGFP-INPP5E variants were coexpressed in HEK293T cells with TULP3-myc as indicated. Lysates were immunoprecipitated with GFP-Trap beads and analyzed by Western blot with TULP3 and EGFP antibodies, as indicated. (b) Quantitation of TULP3-myc co-IP by the indicated EGFP-INPP5E constructs in HEK293T cells. Co-IP levels, expressed as percentage of WT, are normalized by the amounts of both immunoprecipitated EGFP-INPP5Es and TULP3-myc lysate levels. Data are mean ± SEM of n=7,7,4,7,7,4,4 independent experiments and were analyzed by one-way ANOVA followed by Tukey multiple comparisons tests. Significance relative to WT is shown

Journal: eLife

Article Title: Multiple ciliary localization signals control INPP5E ciliary targeting

doi: 10.7554/elife.78383

Figure Lengend Snippet: Figure 7. CLS2 and CLS3 regulate INPP5E binding to TULP3 and CEP164. (a) The indicated EGFP-INPP5E variants were coexpressed in HEK293T cells with TULP3-myc as indicated. Lysates were immunoprecipitated with GFP-Trap beads and analyzed by Western blot with TULP3 and EGFP antibodies, as indicated. (b) Quantitation of TULP3-myc co-IP by the indicated EGFP-INPP5E constructs in HEK293T cells. Co-IP levels, expressed as percentage of WT, are normalized by the amounts of both immunoprecipitated EGFP-INPP5Es and TULP3-myc lysate levels. Data are mean ± SEM of n=7,7,4,7,7,4,4 independent experiments and were analyzed by one-way ANOVA followed by Tukey multiple comparisons tests. Significance relative to WT is shown

Article Snippet: DOI: https://doi.org/10.7554/eLife.78383 27 of 43 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Cell line (Homo sapiens) Puromycin- sensitive hTERT- RPE1 This study Puromycin Acetyltransferase (PAC)KO cells Also used in Gonçalves et al., 2021 Cell line (Homo sapiens) INPP5E- KO hTERTRPE1 This study Clone 3 Derived from puromycin- sensitive hTERT- RPE1 Cell line (Homo sapiens) INPP5E- KO hTERTRPE1 This study Clone 12 Derived from puromycin- sensitive hTERT- RPE1 Cell line (Homo sapiens) 293T ATCC Cat # CRL- 3216 Derived from human embryonic kidney Cell line (Homo sapiens) HeLa ATCC Cat # CCL- 2 Derived from cervical carcinoma Cell line (Homo sapiens) Jurkat, Clone E6- 1 ATCC Cat # TIB- 152 T lymphoblasts from acute T cell leukemia Cell line (Homo sapiens) Raji ATCC Cat # CCL- 86 Lymphoblast- like cells from Burkitt’s lymphoma Cell line (Mus musculus) Csnk2a1- WT MEFs Loukil et al., 2021 N/A Control mouse embryonic fibroblasts (MEFs) Cell line (Mus musculus) Csnk2a1- KO MEFs Loukil et al., 2021 N/A Casein kinase 2 subunit alpha- null MEFs Antibody Anti- acetylated α-tubulin (mouse monoclonal) Sigma- Aldrich (Merck) Cat # T7451 (clone 6- 11B- 1) IF: 1:10,000 Antibody Anti-α-tubulin (mouse monoclonal) Proteintech Cat # 66031–1- Ig WB: 1:1000 Antibody Anti-γ-tubulin (mouse monoclonal) Santa Cruz Cat # sc- 17787 IF: 1:200 Antibody Anti- EGFP (mouse monoclonal) Proteintech Cat # 66002–1- Ig WB: 1:1000 Antibody Anti- EGFP (mouse monoclonal) Santa Cruz Cat # sc- 9996 IF: 1:200 Antibody Anti- ARL13B (mouse monoclonal) Proteintech Cat # 66739–1- Ig WB: 1:1000 Antibody Anti- Flag (mouse monoclonal) Sigma- Aldrich (Merck) Cat # F3165 (clone M2) WB: 1:2000 Antibody Anti- polyglutamylated tubulin (mouse monoclonal) Adipogen Cat # GT335 IF: 1:2000 Antibody Anti- EGFP (rabbit polyclonal) Proteintech Cat # 50430–2- AP IF: 1:200 WB: 1:1000 Antibody Anti- RPGR (rabbit polyclonal) Proteintech Cat # 16891–1- AP WB: 1:1000 Antibody Anti- TULP3 (rabbit polyclonal) Proteintech Cat # 13637–1- AP IF: 1:750 WB: 1:2000 Antibody Anti- CEP164 (rabbit polyclonal) Proteintech Cat # 22227–1- AP WB: 1:1000 Antibody Anti- ATG16L (rabbit polyclonal) MBL Cat # PM040 WB: 1:1000 Continued on next page Continued Cilleros- Rodriguez, Martin- Morales et al. eLife 2022;11:e78383.

Techniques: Binding Assay, Immunoprecipitation, Western Blot, Quantitation Assay, Co-Immunoprecipitation Assay, Construct

Figure 8. CLS1 and CLS4 jointly modulate the ATG16L1-INPP5E interaction. (a) The indicated EGFP-INPP5E constructs were coexpressed in HEK293T cells with Flag-ATG16L1, as shown. Lysates were immunoprecipitated with GFP-Trap beads and analyzed by Western blot with the indicated antibodies. (b) Quantitation of Flag-ATG16L1 co-IP by the indicated EGFP-INPP5E constructs in HEK293T cells. Co-IP levels, expressed as percentage of WT, are normalized by the amounts of both immunoprecipitated EGFP-INPP5Es and Flag-ATG16L1 lysate levels. Data are mean ± SEM of n=3 independent experiments and were analyzed by one-way ANOVA followed by Dunnett tests relative to WT. Significance is shown as p<0.01(**). (c) Schema of INPP5E structure depicting CLS1-4 and the proteins through which they regulate INPP5E ciliary targeting, as shown herein.

Journal: eLife

Article Title: Multiple ciliary localization signals control INPP5E ciliary targeting

doi: 10.7554/elife.78383

Figure Lengend Snippet: Figure 8. CLS1 and CLS4 jointly modulate the ATG16L1-INPP5E interaction. (a) The indicated EGFP-INPP5E constructs were coexpressed in HEK293T cells with Flag-ATG16L1, as shown. Lysates were immunoprecipitated with GFP-Trap beads and analyzed by Western blot with the indicated antibodies. (b) Quantitation of Flag-ATG16L1 co-IP by the indicated EGFP-INPP5E constructs in HEK293T cells. Co-IP levels, expressed as percentage of WT, are normalized by the amounts of both immunoprecipitated EGFP-INPP5Es and Flag-ATG16L1 lysate levels. Data are mean ± SEM of n=3 independent experiments and were analyzed by one-way ANOVA followed by Dunnett tests relative to WT. Significance is shown as p<0.01(**). (c) Schema of INPP5E structure depicting CLS1-4 and the proteins through which they regulate INPP5E ciliary targeting, as shown herein.

Article Snippet: DOI: https://doi.org/10.7554/eLife.78383 27 of 43 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Cell line (Homo sapiens) Puromycin- sensitive hTERT- RPE1 This study Puromycin Acetyltransferase (PAC)KO cells Also used in Gonçalves et al., 2021 Cell line (Homo sapiens) INPP5E- KO hTERTRPE1 This study Clone 3 Derived from puromycin- sensitive hTERT- RPE1 Cell line (Homo sapiens) INPP5E- KO hTERTRPE1 This study Clone 12 Derived from puromycin- sensitive hTERT- RPE1 Cell line (Homo sapiens) 293T ATCC Cat # CRL- 3216 Derived from human embryonic kidney Cell line (Homo sapiens) HeLa ATCC Cat # CCL- 2 Derived from cervical carcinoma Cell line (Homo sapiens) Jurkat, Clone E6- 1 ATCC Cat # TIB- 152 T lymphoblasts from acute T cell leukemia Cell line (Homo sapiens) Raji ATCC Cat # CCL- 86 Lymphoblast- like cells from Burkitt’s lymphoma Cell line (Mus musculus) Csnk2a1- WT MEFs Loukil et al., 2021 N/A Control mouse embryonic fibroblasts (MEFs) Cell line (Mus musculus) Csnk2a1- KO MEFs Loukil et al., 2021 N/A Casein kinase 2 subunit alpha- null MEFs Antibody Anti- acetylated α-tubulin (mouse monoclonal) Sigma- Aldrich (Merck) Cat # T7451 (clone 6- 11B- 1) IF: 1:10,000 Antibody Anti-α-tubulin (mouse monoclonal) Proteintech Cat # 66031–1- Ig WB: 1:1000 Antibody Anti-γ-tubulin (mouse monoclonal) Santa Cruz Cat # sc- 17787 IF: 1:200 Antibody Anti- EGFP (mouse monoclonal) Proteintech Cat # 66002–1- Ig WB: 1:1000 Antibody Anti- EGFP (mouse monoclonal) Santa Cruz Cat # sc- 9996 IF: 1:200 Antibody Anti- ARL13B (mouse monoclonal) Proteintech Cat # 66739–1- Ig WB: 1:1000 Antibody Anti- Flag (mouse monoclonal) Sigma- Aldrich (Merck) Cat # F3165 (clone M2) WB: 1:2000 Antibody Anti- polyglutamylated tubulin (mouse monoclonal) Adipogen Cat # GT335 IF: 1:2000 Antibody Anti- EGFP (rabbit polyclonal) Proteintech Cat # 50430–2- AP IF: 1:200 WB: 1:1000 Antibody Anti- RPGR (rabbit polyclonal) Proteintech Cat # 16891–1- AP WB: 1:1000 Antibody Anti- TULP3 (rabbit polyclonal) Proteintech Cat # 13637–1- AP IF: 1:750 WB: 1:2000 Antibody Anti- CEP164 (rabbit polyclonal) Proteintech Cat # 22227–1- AP WB: 1:1000 Antibody Anti- ATG16L (rabbit polyclonal) MBL Cat # PM040 WB: 1:1000 Continued on next page Continued Cilleros- Rodriguez, Martin- Morales et al. eLife 2022;11:e78383.

Techniques: Construct, Immunoprecipitation, Western Blot, Quantitation Assay, Co-Immunoprecipitation Assay