nbd-ps (16:0–12:0 Search Results


popc  (Croda International Plc)
90
Croda International Plc popc
Translocon reconstitution and fluorescence assays. A) Sucrose gradient purification of detergent-solubilized ribosome/Sec61 translocon complexes. A 260 UV trace (top), total protein staining (middle), and Western blot analysis (bottom) of the sucrose gradient fractions. HEK293T and sheep whole pancreatic lysates are included as antibody controls. The fraction used in the reconstitution (red) is highlighted together with the composition of <t>the</t> <t>LUVs</t> (blue). The uncropped gels are available in Figure S1 in the SI . B) Chemical structures of the lipids used in experiments. The LUVs are made of <t>POPC</t> with 0.4% (mol) of sn -2 acyl chain-NBD-labeled PC (NBD PC) or PS (NBD PS). C) The sodium dithionite, added to the supernatant, chemically reduces the fluorescent NBD PC or NBD PS to nonfluorescent ABD PC or ABD PS in the outer leaflet. This leads to a decrease of fluorescence intensity by ≈50% of its initial value. In the presence of a scramblase, all NBD-labeled lipids eventually reach the outer leaflet resulting in an almost complete loss of NBD fluorescence. D) The addition of bovine serum albumin (BSA) to the sample leads to partial quenching (≈50%) of NBD fluorescence. Hence, in the absence (presence) of a scramblase, a decrease of ≈25% (≈50%) is expected. E) Results from the dithionite assay. Fluorescence before dithionite addition is normalized to 1, and the curves show a mean of N = 2 repeats. Dashed line shows the expected result in the case of no scrambling (0.50), and further decrease is likely due to the dimming of NBD over time. In the absence of proteins, the NBD PC and NBD PS in the outer leaflet is chemically reduced, resulting in a fluorescence intensity decrease to ≈50% of the original value. In the presence of proteins, the value further decreases to ≈15%, i.e. the majority of the NBD-labeled lipids originally in the inner leaflet are scrambled to the outer leaflet and reduced therein. The presence of Sec61 inhibitors (here either Ipomoeassin F, “IpomF” or Apratoxin A, “AprA”) do not affect scrambling. Overall, the interaction between NBD PS and dithionite seems slower, likely due to the electrostatic repulsion between anionic PS head groups and dithionite. F) Results from BSA assay, confirming the findings of panel E). The dashed lines shows the expected results in the case there is (0.50) or is not (0.75) scrambling. The curves show mean of N = 2 repeats. The presence of Sec61 inhibitors (IpomF or AprA) had no effect on scrambling. Some graphical elements in panels C and D were created with BioRender.com, and they are available online: Šimek, J. (2025) https://BioRender.com/sx9c95m .
Popc, supplied by Croda International Plc, 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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popc - by Bioz Stars, 2026-10
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ps(16:0/18:1  (Avanti Inc)
90
Avanti Inc ps(16:0/18:1
Translocon reconstitution and fluorescence assays. A) Sucrose gradient purification of detergent-solubilized ribosome/Sec61 translocon complexes. A 260 UV trace (top), total protein staining (middle), and Western blot analysis (bottom) of the sucrose gradient fractions. HEK293T and sheep whole pancreatic lysates are included as antibody controls. The fraction used in the reconstitution (red) is highlighted together with the composition of <t>the</t> <t>LUVs</t> (blue). The uncropped gels are available in Figure S1 in the SI . B) Chemical structures of the lipids used in experiments. The LUVs are made of <t>POPC</t> with 0.4% (mol) of sn -2 acyl chain-NBD-labeled PC (NBD PC) or PS (NBD PS). C) The sodium dithionite, added to the supernatant, chemically reduces the fluorescent NBD PC or NBD PS to nonfluorescent ABD PC or ABD PS in the outer leaflet. This leads to a decrease of fluorescence intensity by ≈50% of its initial value. In the presence of a scramblase, all NBD-labeled lipids eventually reach the outer leaflet resulting in an almost complete loss of NBD fluorescence. D) The addition of bovine serum albumin (BSA) to the sample leads to partial quenching (≈50%) of NBD fluorescence. Hence, in the absence (presence) of a scramblase, a decrease of ≈25% (≈50%) is expected. E) Results from the dithionite assay. Fluorescence before dithionite addition is normalized to 1, and the curves show a mean of N = 2 repeats. Dashed line shows the expected result in the case of no scrambling (0.50), and further decrease is likely due to the dimming of NBD over time. In the absence of proteins, the NBD PC and NBD PS in the outer leaflet is chemically reduced, resulting in a fluorescence intensity decrease to ≈50% of the original value. In the presence of proteins, the value further decreases to ≈15%, i.e. the majority of the NBD-labeled lipids originally in the inner leaflet are scrambled to the outer leaflet and reduced therein. The presence of Sec61 inhibitors (here either Ipomoeassin F, “IpomF” or Apratoxin A, “AprA”) do not affect scrambling. Overall, the interaction between NBD PS and dithionite seems slower, likely due to the electrostatic repulsion between anionic PS head groups and dithionite. F) Results from BSA assay, confirming the findings of panel E). The dashed lines shows the expected results in the case there is (0.50) or is not (0.75) scrambling. The curves show mean of N = 2 repeats. The presence of Sec61 inhibitors (IpomF or AprA) had no effect on scrambling. Some graphical elements in panels C and D were created with BioRender.com, and they are available online: Šimek, J. (2025) https://BioRender.com/sx9c95m .
Ps(16:0/18:1, supplied by Avanti Inc, 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/product/nbd-ps+(16%3A0%E2%80%9312%3A0/nbd+ps++16+0+12+0/pmc08590225-109-2-6
Average 90 stars, based on 1 article reviews
ps(16:0/18:1 - by Bioz Stars, 2026-10
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16 0 12 0 n nbd ps  (Avanti Polar)
90
Avanti Polar 16 0 12 0 n nbd ps
Translocon reconstitution and fluorescence assays. A) Sucrose gradient purification of detergent-solubilized ribosome/Sec61 translocon complexes. A 260 UV trace (top), total protein staining (middle), and Western blot analysis (bottom) of the sucrose gradient fractions. HEK293T and sheep whole pancreatic lysates are included as antibody controls. The fraction used in the reconstitution (red) is highlighted together with the composition of <t>the</t> <t>LUVs</t> (blue). The uncropped gels are available in Figure S1 in the SI . B) Chemical structures of the lipids used in experiments. The LUVs are made of <t>POPC</t> with 0.4% (mol) of sn -2 acyl chain-NBD-labeled PC (NBD PC) or PS (NBD PS). C) The sodium dithionite, added to the supernatant, chemically reduces the fluorescent NBD PC or NBD PS to nonfluorescent ABD PC or ABD PS in the outer leaflet. This leads to a decrease of fluorescence intensity by ≈50% of its initial value. In the presence of a scramblase, all NBD-labeled lipids eventually reach the outer leaflet resulting in an almost complete loss of NBD fluorescence. D) The addition of bovine serum albumin (BSA) to the sample leads to partial quenching (≈50%) of NBD fluorescence. Hence, in the absence (presence) of a scramblase, a decrease of ≈25% (≈50%) is expected. E) Results from the dithionite assay. Fluorescence before dithionite addition is normalized to 1, and the curves show a mean of N = 2 repeats. Dashed line shows the expected result in the case of no scrambling (0.50), and further decrease is likely due to the dimming of NBD over time. In the absence of proteins, the NBD PC and NBD PS in the outer leaflet is chemically reduced, resulting in a fluorescence intensity decrease to ≈50% of the original value. In the presence of proteins, the value further decreases to ≈15%, i.e. the majority of the NBD-labeled lipids originally in the inner leaflet are scrambled to the outer leaflet and reduced therein. The presence of Sec61 inhibitors (here either Ipomoeassin F, “IpomF” or Apratoxin A, “AprA”) do not affect scrambling. Overall, the interaction between NBD PS and dithionite seems slower, likely due to the electrostatic repulsion between anionic PS head groups and dithionite. F) Results from BSA assay, confirming the findings of panel E). The dashed lines shows the expected results in the case there is (0.50) or is not (0.75) scrambling. The curves show mean of N = 2 repeats. The presence of Sec61 inhibitors (IpomF or AprA) had no effect on scrambling. Some graphical elements in panels C and D were created with BioRender.com, and they are available online: Šimek, J. (2025) https://BioRender.com/sx9c95m .
16 0 12 0 N Nbd Ps, supplied by Avanti Polar, 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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16 0 12 0 n nbd ps - by Bioz Stars, 2026-10
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16:0-12:0 nbd ps  (BOC Sciences)
N/A
NBD PS is a fluorophore having fluorescent 7-nitro-2-1,3-benzoxadiazol-4-yl (NBD) label attached to PS. Phosphatidylserine (PS) is involved in the regulation of many enzymes and structural components. It serves as an essential factor in the clearance
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Translocon reconstitution and fluorescence assays. A) Sucrose gradient purification of detergent-solubilized ribosome/Sec61 translocon complexes. A 260 UV trace (top), total protein staining (middle), and Western blot analysis (bottom) of the sucrose gradient fractions. HEK293T and sheep whole pancreatic lysates are included as antibody controls. The fraction used in the reconstitution (red) is highlighted together with the composition of the LUVs (blue). The uncropped gels are available in Figure S1 in the SI . B) Chemical structures of the lipids used in experiments. The LUVs are made of POPC with 0.4% (mol) of sn -2 acyl chain-NBD-labeled PC (NBD PC) or PS (NBD PS). C) The sodium dithionite, added to the supernatant, chemically reduces the fluorescent NBD PC or NBD PS to nonfluorescent ABD PC or ABD PS in the outer leaflet. This leads to a decrease of fluorescence intensity by ≈50% of its initial value. In the presence of a scramblase, all NBD-labeled lipids eventually reach the outer leaflet resulting in an almost complete loss of NBD fluorescence. D) The addition of bovine serum albumin (BSA) to the sample leads to partial quenching (≈50%) of NBD fluorescence. Hence, in the absence (presence) of a scramblase, a decrease of ≈25% (≈50%) is expected. E) Results from the dithionite assay. Fluorescence before dithionite addition is normalized to 1, and the curves show a mean of N = 2 repeats. Dashed line shows the expected result in the case of no scrambling (0.50), and further decrease is likely due to the dimming of NBD over time. In the absence of proteins, the NBD PC and NBD PS in the outer leaflet is chemically reduced, resulting in a fluorescence intensity decrease to ≈50% of the original value. In the presence of proteins, the value further decreases to ≈15%, i.e. the majority of the NBD-labeled lipids originally in the inner leaflet are scrambled to the outer leaflet and reduced therein. The presence of Sec61 inhibitors (here either Ipomoeassin F, “IpomF” or Apratoxin A, “AprA”) do not affect scrambling. Overall, the interaction between NBD PS and dithionite seems slower, likely due to the electrostatic repulsion between anionic PS head groups and dithionite. F) Results from BSA assay, confirming the findings of panel E). The dashed lines shows the expected results in the case there is (0.50) or is not (0.75) scrambling. The curves show mean of N = 2 repeats. The presence of Sec61 inhibitors (IpomF or AprA) had no effect on scrambling. Some graphical elements in panels C and D were created with BioRender.com, and they are available online: Šimek, J. (2025) https://BioRender.com/sx9c95m .

Journal: Journal of the American Chemical Society

Article Title: Lipid Scrambling Pathways in the Sec61 Translocon Complex

doi: 10.1021/jacs.4c11142

Figure Lengend Snippet: Translocon reconstitution and fluorescence assays. A) Sucrose gradient purification of detergent-solubilized ribosome/Sec61 translocon complexes. A 260 UV trace (top), total protein staining (middle), and Western blot analysis (bottom) of the sucrose gradient fractions. HEK293T and sheep whole pancreatic lysates are included as antibody controls. The fraction used in the reconstitution (red) is highlighted together with the composition of the LUVs (blue). The uncropped gels are available in Figure S1 in the SI . B) Chemical structures of the lipids used in experiments. The LUVs are made of POPC with 0.4% (mol) of sn -2 acyl chain-NBD-labeled PC (NBD PC) or PS (NBD PS). C) The sodium dithionite, added to the supernatant, chemically reduces the fluorescent NBD PC or NBD PS to nonfluorescent ABD PC or ABD PS in the outer leaflet. This leads to a decrease of fluorescence intensity by ≈50% of its initial value. In the presence of a scramblase, all NBD-labeled lipids eventually reach the outer leaflet resulting in an almost complete loss of NBD fluorescence. D) The addition of bovine serum albumin (BSA) to the sample leads to partial quenching (≈50%) of NBD fluorescence. Hence, in the absence (presence) of a scramblase, a decrease of ≈25% (≈50%) is expected. E) Results from the dithionite assay. Fluorescence before dithionite addition is normalized to 1, and the curves show a mean of N = 2 repeats. Dashed line shows the expected result in the case of no scrambling (0.50), and further decrease is likely due to the dimming of NBD over time. In the absence of proteins, the NBD PC and NBD PS in the outer leaflet is chemically reduced, resulting in a fluorescence intensity decrease to ≈50% of the original value. In the presence of proteins, the value further decreases to ≈15%, i.e. the majority of the NBD-labeled lipids originally in the inner leaflet are scrambled to the outer leaflet and reduced therein. The presence of Sec61 inhibitors (here either Ipomoeassin F, “IpomF” or Apratoxin A, “AprA”) do not affect scrambling. Overall, the interaction between NBD PS and dithionite seems slower, likely due to the electrostatic repulsion between anionic PS head groups and dithionite. F) Results from BSA assay, confirming the findings of panel E). The dashed lines shows the expected results in the case there is (0.50) or is not (0.75) scrambling. The curves show mean of N = 2 repeats. The presence of Sec61 inhibitors (IpomF or AprA) had no effect on scrambling. Some graphical elements in panels C and D were created with BioRender.com, and they are available online: Šimek, J. (2025) https://BioRender.com/sx9c95m .

Article Snippet: Large Unilamellar Vesicles (LUVs), composed of 99.6% (mol) POPC and 0.4% (mol) of 16:0–12:0 NBD PS; or 0.4% (mol) of 16:0–06:0 NBD PC (Avanti), were prepared by extrusion.

Techniques: Fluorescence, Purification, Staining, Western Blot, Labeling