gfp sec61β 121159 Search Results


plasmid gfp sec61β  (Addgene inc)
93
Addgene inc plasmid gfp sec61β
In vivo scrambling activity assay showing TMEM41B shuttles PC between the ER membrane leaflets (A) Schematic of metabolic labeling and click-chemistry to detect PC in Huh7 TMEM41B WT/KO cells. Cells are incubated with alkyne-choline for synthesis into alkyne-PC at the ER outer leaflet. In wild-type cells, alkyne-PC will shuttle across the bilayer into the inner leaflet of the ER. Digitonin can permeabilizing the cell surface membrane, but the endo-membrane would be intact. Therefore, the click-chemistry reagents could only label alkyne-PC on the outer leaflet of the ER but could not access the inner leaflet of ER membrane. The absence of scramblases in the ER of mutant cells would trap more alkyne-PC on the outer leaflet and increase the fluorescent signal. (B) Representative western blots for validation of TMEM41B deficiency in Huh7 cells. (C) Representative confocal images showing accumulation of alkyne-PC signal on the outer leaflet of the ER in the absence of TMEM41B (n=3 independent experiments). CRISPR/Cas9-mediated control or TMEM41B KO Huh7 cells incubating with alkyne-choline are clicked with 5-TAMRA azide and visualized by confocal microscopy. The ER is marked by <t>GFP-SEC61β.</t> The middle image is a zoomed in image from the white box in the left image. Right: surface plots depicting alkyne-PC signals. Scale bars, 5 μm. (D) Quantification of colocalization of signals of alkyne-PC and GFP-SEC61β (left) and total fluorescence signals (a.u.) of alkyne-PC (right) from (B). Data are presented as mean ± SEM. n.s., no significance, ∗∗∗∗p < 0.0001 (two-tailed Student’s t test).
Plasmid Gfp Sec61β, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/gfp+sec61%CE%B2+121159/GFP-+SEC61B+(Plasmid+%23121159)/pmc09043772-140-0-5
Average 93 stars, based on 1 article reviews
plasmid gfp sec61β - by Bioz Stars, 2026-09
93/100 stars
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sec61β 1 96aa  (Addgene inc)
94
Addgene inc sec61β 1 96aa
In vivo scrambling activity assay showing TMEM41B shuttles PC between the ER membrane leaflets (A) Schematic of metabolic labeling and click-chemistry to detect PC in Huh7 TMEM41B WT/KO cells. Cells are incubated with alkyne-choline for synthesis into alkyne-PC at the ER outer leaflet. In wild-type cells, alkyne-PC will shuttle across the bilayer into the inner leaflet of the ER. Digitonin can permeabilizing the cell surface membrane, but the endo-membrane would be intact. Therefore, the click-chemistry reagents could only label alkyne-PC on the outer leaflet of the ER but could not access the inner leaflet of ER membrane. The absence of scramblases in the ER of mutant cells would trap more alkyne-PC on the outer leaflet and increase the fluorescent signal. (B) Representative western blots for validation of TMEM41B deficiency in Huh7 cells. (C) Representative confocal images showing accumulation of alkyne-PC signal on the outer leaflet of the ER in the absence of TMEM41B (n=3 independent experiments). CRISPR/Cas9-mediated control or TMEM41B KO Huh7 cells incubating with alkyne-choline are clicked with 5-TAMRA azide and visualized by confocal microscopy. The ER is marked by <t>GFP-SEC61β.</t> The middle image is a zoomed in image from the white box in the left image. Right: surface plots depicting alkyne-PC signals. Scale bars, 5 μm. (D) Quantification of colocalization of signals of alkyne-PC and GFP-SEC61β (left) and total fluorescence signals (a.u.) of alkyne-PC (right) from (B). Data are presented as mean ± SEM. n.s., no significance, ∗∗∗∗p < 0.0001 (two-tailed Student’s t test).
Sec61β 1 96aa, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/gfp+sec61%CE%B2+121159/RA-Sec61%CE%B2+(Plasmid+%23153978)/pmc08351558__mbc___32___1147___s001-34-300-320
Average 94 stars, based on 1 article reviews
sec61β 1 96aa - by Bioz Stars, 2026-09
94/100 stars
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mctp2 rhd 640 878aa  (Addgene inc)
93
Addgene inc mctp2 rhd 640 878aa
In vivo scrambling activity assay showing TMEM41B shuttles PC between the ER membrane leaflets (A) Schematic of metabolic labeling and click-chemistry to detect PC in Huh7 TMEM41B WT/KO cells. Cells are incubated with alkyne-choline for synthesis into alkyne-PC at the ER outer leaflet. In wild-type cells, alkyne-PC will shuttle across the bilayer into the inner leaflet of the ER. Digitonin can permeabilizing the cell surface membrane, but the endo-membrane would be intact. Therefore, the click-chemistry reagents could only label alkyne-PC on the outer leaflet of the ER but could not access the inner leaflet of ER membrane. The absence of scramblases in the ER of mutant cells would trap more alkyne-PC on the outer leaflet and increase the fluorescent signal. (B) Representative western blots for validation of TMEM41B deficiency in Huh7 cells. (C) Representative confocal images showing accumulation of alkyne-PC signal on the outer leaflet of the ER in the absence of TMEM41B (n=3 independent experiments). CRISPR/Cas9-mediated control or TMEM41B KO Huh7 cells incubating with alkyne-choline are clicked with 5-TAMRA azide and visualized by confocal microscopy. The ER is marked by <t>GFP-SEC61β.</t> The middle image is a zoomed in image from the white box in the left image. Right: surface plots depicting alkyne-PC signals. Scale bars, 5 μm. (D) Quantification of colocalization of signals of alkyne-PC and GFP-SEC61β (left) and total fluorescence signals (a.u.) of alkyne-PC (right) from (B). Data are presented as mean ± SEM. n.s., no significance, ∗∗∗∗p < 0.0001 (two-tailed Student’s t test).
Mctp2 Rhd 640 878aa, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/gfp+sec61%CE%B2+121159/mCh-Rab5+(Plasmid+%2349201)/pmc08351558__mbc___32___1147___s001-34-312-320
Average 93 stars, based on 1 article reviews
mctp2 rhd 640 878aa - by Bioz Stars, 2026-09
93/100 stars
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name region source reference mctp1 gfp full length  (Addgene inc)
93
Addgene inc name region source reference mctp1 gfp full length
In vivo scrambling activity assay showing TMEM41B shuttles PC between the ER membrane leaflets (A) Schematic of metabolic labeling and click-chemistry to detect PC in Huh7 TMEM41B WT/KO cells. Cells are incubated with alkyne-choline for synthesis into alkyne-PC at the ER outer leaflet. In wild-type cells, alkyne-PC will shuttle across the bilayer into the inner leaflet of the ER. Digitonin can permeabilizing the cell surface membrane, but the endo-membrane would be intact. Therefore, the click-chemistry reagents could only label alkyne-PC on the outer leaflet of the ER but could not access the inner leaflet of ER membrane. The absence of scramblases in the ER of mutant cells would trap more alkyne-PC on the outer leaflet and increase the fluorescent signal. (B) Representative western blots for validation of TMEM41B deficiency in Huh7 cells. (C) Representative confocal images showing accumulation of alkyne-PC signal on the outer leaflet of the ER in the absence of TMEM41B (n=3 independent experiments). CRISPR/Cas9-mediated control or TMEM41B KO Huh7 cells incubating with alkyne-choline are clicked with 5-TAMRA azide and visualized by confocal microscopy. The ER is marked by <t>GFP-SEC61β.</t> The middle image is a zoomed in image from the white box in the left image. Right: surface plots depicting alkyne-PC signals. Scale bars, 5 μm. (D) Quantification of colocalization of signals of alkyne-PC and GFP-SEC61β (left) and total fluorescence signals (a.u.) of alkyne-PC (right) from (B). Data are presented as mean ± SEM. n.s., no significance, ∗∗∗∗p < 0.0001 (two-tailed Student’s t test).
Name Region Source Reference Mctp1 Gfp Full Length, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/gfp+sec61%CE%B2+121159/mCh-Climp63+(Plasmid+%23136293)/pmc08351558__mbc___32___1147___s001-34-191-218
Average 93 stars, based on 1 article reviews
name region source reference mctp1 gfp full length - by Bioz Stars, 2026-09
93/100 stars
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Image Search Results


In vivo scrambling activity assay showing TMEM41B shuttles PC between the ER membrane leaflets (A) Schematic of metabolic labeling and click-chemistry to detect PC in Huh7 TMEM41B WT/KO cells. Cells are incubated with alkyne-choline for synthesis into alkyne-PC at the ER outer leaflet. In wild-type cells, alkyne-PC will shuttle across the bilayer into the inner leaflet of the ER. Digitonin can permeabilizing the cell surface membrane, but the endo-membrane would be intact. Therefore, the click-chemistry reagents could only label alkyne-PC on the outer leaflet of the ER but could not access the inner leaflet of ER membrane. The absence of scramblases in the ER of mutant cells would trap more alkyne-PC on the outer leaflet and increase the fluorescent signal. (B) Representative western blots for validation of TMEM41B deficiency in Huh7 cells. (C) Representative confocal images showing accumulation of alkyne-PC signal on the outer leaflet of the ER in the absence of TMEM41B (n=3 independent experiments). CRISPR/Cas9-mediated control or TMEM41B KO Huh7 cells incubating with alkyne-choline are clicked with 5-TAMRA azide and visualized by confocal microscopy. The ER is marked by GFP-SEC61β. The middle image is a zoomed in image from the white box in the left image. Right: surface plots depicting alkyne-PC signals. Scale bars, 5 μm. (D) Quantification of colocalization of signals of alkyne-PC and GFP-SEC61β (left) and total fluorescence signals (a.u.) of alkyne-PC (right) from (B). Data are presented as mean ± SEM. n.s., no significance, ∗∗∗∗p < 0.0001 (two-tailed Student’s t test).

Journal: STAR Protocols

Article Title: In vitro and in vivo assay of the ER lipid scramblase TMEM41B

doi: 10.1016/j.xpro.2022.101333

Figure Lengend Snippet: In vivo scrambling activity assay showing TMEM41B shuttles PC between the ER membrane leaflets (A) Schematic of metabolic labeling and click-chemistry to detect PC in Huh7 TMEM41B WT/KO cells. Cells are incubated with alkyne-choline for synthesis into alkyne-PC at the ER outer leaflet. In wild-type cells, alkyne-PC will shuttle across the bilayer into the inner leaflet of the ER. Digitonin can permeabilizing the cell surface membrane, but the endo-membrane would be intact. Therefore, the click-chemistry reagents could only label alkyne-PC on the outer leaflet of the ER but could not access the inner leaflet of ER membrane. The absence of scramblases in the ER of mutant cells would trap more alkyne-PC on the outer leaflet and increase the fluorescent signal. (B) Representative western blots for validation of TMEM41B deficiency in Huh7 cells. (C) Representative confocal images showing accumulation of alkyne-PC signal on the outer leaflet of the ER in the absence of TMEM41B (n=3 independent experiments). CRISPR/Cas9-mediated control or TMEM41B KO Huh7 cells incubating with alkyne-choline are clicked with 5-TAMRA azide and visualized by confocal microscopy. The ER is marked by GFP-SEC61β. The middle image is a zoomed in image from the white box in the left image. Right: surface plots depicting alkyne-PC signals. Scale bars, 5 μm. (D) Quantification of colocalization of signals of alkyne-PC and GFP-SEC61β (left) and total fluorescence signals (a.u.) of alkyne-PC (right) from (B). Data are presented as mean ± SEM. n.s., no significance, ∗∗∗∗p < 0.0001 (two-tailed Student’s t test).

Article Snippet: Plasmid: GFP- SEC61β , , Addgene Cat# 121159.

Techniques: In Vivo, Activity Assay, Membrane, Labeling, Incubation, Mutagenesis, Western Blot, Biomarker Discovery, CRISPR, Control, Confocal Microscopy, Fluorescence, Two Tailed Test

Journal: STAR Protocols

Article Title: In vitro and in vivo assay of the ER lipid scramblase TMEM41B

doi: 10.1016/j.xpro.2022.101333

Figure Lengend Snippet:

Article Snippet: Plasmid: GFP- SEC61β , , Addgene Cat# 121159.

Techniques: Virus, Recombinant, Protease Inhibitor, Gel Extraction, Plasmid Preparation, Software, Cell Culture, Molecular Weight, Membrane, Blocking Assay, Spectrophotometry, Evaporation