non tagged optn (OriGene)
OriGene is a verified supplier 
OriGene manufactures this product 
92
Structured Review
OriGene
non tagged optn
Non Tagged Optn, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/non+tagged+optn/Optineurin+(OPTN)+(NM_001008211)+Human+Untagged+Clone/bio_rxiv__2024__05__31__596776-116-49-52
Average 92 stars, based on 1 article reviews
Non Tagged Optn, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/non+tagged+optn/Optineurin+(OPTN)+(NM_001008211)+Human+Untagged+Clone/bio_rxiv__2024__05__31__596776-116-49-52
Average 92 stars, based on 1 article reviews
non tagged optn - by Bioz Stars,
2026-09
92/100 stars
Images
1) Product Images from "PEX14 acts as the interface linking optineurin to cell type-selective pexophagy"
Article Title: PEX14 acts as the interface linking optineurin to cell type-selective pexophagy
Journal: bioRxiv
doi: 10.1101/2024.05.31.596776
Figure Legend Snippet: HCT116 and HeLa cells stably expressing po-mKeima were transfected with a plasmid encoding GFP or OPTN-GFP and cultured as described in the Materials and Methods section. After two days, the cells were harvested and processed for FACS analysis. To measure the percentage of cells undergoing pexophagy, single-cell GFP-positive populations were gated for a decrease in fluorescence intensity in the neutral channel. a , Representative flow cytometry plots of each group (n = 3). The different colors represent the cell density at a given position. b , Quantification of the percentage of cells in the gated area. The data are shown as the mean ± SD and represent the values of 3 independent biological replicates. All conditions were statistically compared to the GFP condition (ns, non-significant; **, p < 0.01).
Techniques Used: Stable Transfection, Expressing, Transfection, Plasmid Preparation, Cell Culture, Fluorescence, Flow Cytometry
Figure Legend Snippet: a , Distribution patterns of OPTN-GFP and mCherry-PTS1 in four distinct cell lines, either co-expressing (right panels) or not (left panels) a high-affinity plasma membrane-targeted anti-GFP nanobody (GFP Nb -PM). Before analysis, the cells were incubated for 2 h with 200 nM BafA1. Representative images are shown. The white arrowheads indicate colocalization of OPTN-GFP and mCherry-PTS1. The yellow arrowheads indicate positions where peroxisomes are associated with the plasma membrane. b , Enlargements of the boxed areas in panel a. c , Distribution patterns of endogenous PEX14 and OPTN in HEK-293 and HCT116 cells. d , High-resolution imaging and volume rendering of two mCherry-PTS1/OPTN-GFP aggregates in HEK-293 cells. Scale bars: a-c , 10 μm; d , 500 nm.
Techniques Used: Expressing, Clinical Proteomics, Membrane, Incubation, Imaging
Figure Legend Snippet: a , Schematic representation of the distinct domains and major interactions of the miniTurbo-OPTN-GFP fusion protein. ATG9A, autophagy-related protein 9A; CC, coiled-coil; CYLD, cylindromatosis; HLH, helix-loop-helix; HTT, huntingtin; LZ, leucine zipper domain; LC3/GABARAP, microtubule-associated protein 1A/1B-light chain 3/γ-aminobutyric acid receptor-associated protein; LIR, LC3-interacting region; Rab, Ras-related protein; TBC1D17, TBC1 domain family member 17; TBK1, TANK (TRAF-associated NF-kB activator) binding kinase 1; UBD, ubiquitin-binding domain; ZF, zinc finger. b , Volcano plot displaying the Log 2 -fold changes (FC) in protein abundance alongside the corresponding Log 10 -fold adjusted p-values (pAdj) for 2856 proteins obtained through streptavidin-affinity purification and identification/quantification by LC–MS/MS. The analysis was performed on miniTurbo-OPTN-GFP-expressing HEK-293 cells (n = 4 biological replicates), assayed under conditions with or without 50 µM biotin. Green, blue, and red dots represent miniTurbo-OPTN-GFP, known OPTN interaction partners, and bona fide peroxisomal proteins that could be identified in the affinity-purified fractions. Vertical and horizontal dotted lines denote the FC (2x) and pAdj (0.05) cut-offs, respectively.
Techniques Used: Binding Assay, Ubiquitin Proteomics, Quantitative Proteomics, Affinity Purification, Liquid Chromatography with Mass Spectroscopy, Expressing
Figure Legend Snippet: a , Immunoblot analysis of input and GFP-Trap pull-down fractions from Flp-In T-REx 293 cells expressing the specified proteins. Representative images are shown. Specific protein bands and their degradation products are marked by arrows and arrowheads, respectively. Molecular mass markers (in kDa) are indicated on the left. b , Densitometry quantifications of the relative ratios (RR) of PEX14/GFP and PEX14/OPTN-GFP retained on the GFP-Trap affinity matrix. The total signal intensities of the GFP proteins and PEX14 were both standardized to 100%. The bars represent the mean of 2 biological replicates. The densitometry quantifications of the relative normalized amounts (R-NA) of the proteins under study in the soluble extract input fractions are shown in the . T, transient transfection; F, Flp-In integration; L, lentiviral transduction.
Techniques Used: Western Blot, Expressing, Transfection, Transduction
Figure Legend Snippet: Total cell lysates from lentiviral (L-) transduced Flp-In T-REx 293 cells, which constitutively express either GFP or OPTN-GFP, were subjected to SDS-PAGE followed by immunoblotting using antibodies targeting OPTN and GAPDH. a , A representative image is shown, with specific protein bands indicated by arrows. Note that the 66 kDa band in the OPTN-GFP lane may represent either a degradation product of OPTN-GFP or endogenous OPTN stabilized by overexpression of OPTN-GFP. Molecular mass markers (in kDa) are indicated on the left. b , Densitometry quantifications of the relative normalized amounts (R-NA) of OPTN. The expression levels were normalized to GAPDH, with the amount of OPTN detected in the GFP cells serving as the reference. The amount in the L-OPTN-GFP cells represents the sum of the 66 and 95 kDa bands. The bars represent the mean of 4 biological replicates.
Techniques Used: SDS Page, Western Blot, Over Expression, Expressing
Figure Legend Snippet: Flp-In T-REx 293 cells stably expressing po-mKeima were transfected with a plasmid encoding the indicated GFP-fusion protein and cultured in regular DMEM medium. After two days, the cells were harvested and processed for GFP-Trap and FACS analyses. To measure the percentage of cells undergoing pexophagy, single-cell GFP-positive populations were gated for a decrease in fluorescence intensity in the neutral channel. a , Samples of protein extracts (input) and the GFP Trap were processed for SDS-PAGE followed by immunoblotting using antibodies to GFP, PEX14, and GAPDH. Representative blots are shown. Specific protein bands are marked by arrows. Molecular mass markers (in kDa) are indicated on the left. b , Densitometry quantifications of the relative ratios (RR) of PEX14/GFP and PEX14/OPTN-GFP fusion proteins retained on the GFP-Trap affinity matrix. The total signal intensities of the GFP proteins and PEX14 were both standardized to 100%. The bars represent the mean of 3 biological replicates. The densitometry quantifications of the relative normalized amounts (R-NA) of the proteins under study in the soluble extract input fractions are shown in the . c , Examples of po-mKeima-expressing cells displaying no, weak, moderate, or excessively high levels of pexophagy. The visuals depict overlay images of po-mKeima excited at 440 nm (false color: green) or 586 nm (false color: red). Yellowish (=low 586/440 excitation peak ratio) and reddish (=high 586/440 excitation peak ratio) dots represent peroxisomes and autolysosomes, respectively . Scale bar, 10 µm. d , Representative flow cytometry plots of each group (n = 3). The different colors represent the cell density at a given position. e , Quantification of the percentage of cells in the gated area. The data are shown as the mean ± SD and represent the values of 3 independent biological replicates. All conditions were statistically compared to the OPTN 1-577 -GFP condition (ns, non-significant; *, p < 0.05; **, p < 0.01; ****, p < 0.0001).
Techniques Used: Stable Transfection, Expressing, Transfection, Plasmid Preparation, Cell Culture, Fluorescence, SDS Page, Western Blot, Flow Cytometry
Figure Legend Snippet: Flp-In T-REx 293 cells stably expressing po-mKeima were transfected with a plasmid encoding the indicated GFP-fusion protein and cultured in regular DMEM medium. After two days, the cells were harvested and processed for GFP-Trap and FACS analyses. To measure the percentage of cells undergoing pexophagy, single-cell GFP-positive populations were gated for a decrease in fluorescence intensity in the neutral channel. a , c Samples of protein extracts (input) and the GFP-Trap were processed for SDS-PAGE followed by immunoblotting using antibodies to GFP, PEX14, and GAPDH. Representative blots are shown. Specific protein bands are marked by arrows. Molecular mass markers (in kDa) are indicated on the left. b , d Densitometry quantifications of the relative ratios (RR) of PEX14/GFP and PEX14/OPTN-GFP fusion proteins retained on the GFP-Trap affinity matrix. The total signal intensities of the GFP proteins and PEX14 were both standardized to 100%. The bars represent the mean of 3 biological replicates. The densitometry quantifications of the relative normalized amounts (R-NA) of the proteins under study in the soluble extract input fractions are shown in the . e , Representative flow cytometry plots of each group (n = 3). The different colors represent the cell density at a given position. f , Quantification of the percentage of cells in the gated area. The data are shown as the mean ± SD and represent the values of 3 independent biological replicates. All conditions were statistically compared to the OPTN 1-577 -GFP condition (ns, non-significant; *, p < 0.05; **, p < 0.01; ****, p < 0.0001).
Techniques Used: Stable Transfection, Expressing, Transfection, Plasmid Preparation, Cell Culture, Fluorescence, SDS Page, Western Blot, Flow Cytometry
Figure Legend Snippet: Flp-In T-REx 293 cells stably expressing po-mKeima were transfected with either a control (CT) or OPTN DsiRNA and seeded in 6-well plates. On day 2, the cells were transfected or not with a plasmid encoding OPTN 217-577 -GFP. On day 3, the cells were transfected again with either siRNA CT or siRNA OPTN. On day 4, the cells were harvested and processed for immunoblotting with antisera raised against the indicated proteins. a , Representative blots showing the effectiveness of the OPTN DsiRNA. Specific protein bands are marked by arrows. b , Densitometry quantifications of the relative normalized amounts (R-NA) of the OPTN levels shown in panel a (n = 3). c , Comparison of the migration behavior of endogenous OPTN (CT DsiRNA condition) and OPTN 217-577 -GFP (OPTN DsiRNA / OPTN 217-577 -GFP condition). Molecular mass markers (in kDa) are indicated on the left.
Techniques Used: Stable Transfection, Expressing, Transfection, Control, Plasmid Preparation, Western Blot, Comparison, Migration
Figure Legend Snippet: Flp-In T-REx 293 cells, stably expressing po-mKeima, were transfected with either a control (CT) or OPTN siRNA and seeded in 6-well plates. On day 2, the cells were transfected with either a GFP- or OPTN 217-577 -GFP-encoding plasmid. On day 3, the cells were transfected again with either siRNA CT or siRNA OPTN. On day 4, the cells were harvested and processed for FACS analysis. To measure the percentage of cells undergoing pexophagy, single-cell GFP-positive populations were gated for a decrease in fluorescence intensity in the neutral channel. a , Representative flow cytometry plots of each group (n = 3). The different colors represent the cell density at a given position. b , Quantification of the percentage of cells in the gated area. The data are shown as the mean ± SD and represent the values of 3 independent biological replicates. The impact of OPTN downregulation and OPTN 217-577 -GFP overexpression were statistically compared (ns, non-significant; *, p < 0.05; **, p < 0.01;****, p < 0.0001).
Techniques Used: Stable Transfection, Expressing, Transfection, Control, Plasmid Preparation, Fluorescence, Flow Cytometry, Over Expression
Figure Legend Snippet: Flp-In T-REx 293 cells stably expressing po-mKeima were (co-)transfected with (a) plasmid(s) encoding the indicated protein(s) and cultured in regular DMEM medium. After two days, the cells were harvested and processed for GFP-Trap and FACS analysis. To measure the percentage of cells undergoing pexophagy, single-cell GFP-positive populations were gated for a decrease in fluorescence intensity in the neutral channel. a , Samples of protein extracts (input) and the GFP-Trap were processed for SDS-PAGE followed by immunoblotting using antibodies to GFP, OPTN, and GAPDH. Representative blots are shown. Specific protein bands are marked by arrows. Molecular mass markers (in kDa) are indicated on the left. b , Densitometry quantifications of the relative ratios (RR) of OPTN/GFP and OPTN/PEX14-GFP fusion proteins retained on the GFP-Trap affinity matrix. The total signal intensities of the GFP proteins and PEX14 were both standardized to 100%. The bars represent the mean of 3 biological replicates. The densitometry quantifications of the relative normalized amounts (R-NA) of the proteins under study in the soluble extract input fractions are shown in the . c , e , Representative flow cytometry plots of each group (n = 2-3). The different colors represent the cell density at a given position. d , f , Quantification of the percentage of cells in the gated area. The data are shown as the mean ± SD and represent the values of 2-3 independent biological replicates. Relevant conditions were statistically compared (ns, non-significant; ***, p < 0.001).
Techniques Used: Stable Transfection, Expressing, Transfection, Plasmid Preparation, Cell Culture, Fluorescence, SDS Page, Western Blot, Flow Cytometry
Figure Legend Snippet: a , Structural model of the OPTN-OPTN-PEX14 complex before (left) and after (right) a 50 ns molecular dynamics (MD) simulation, depicted in either cartoon or surface representation. Residues were colored as follows: OPTN 420-510 in green (2x), and PEX14 in red (residues 1-137) and blue (residues 138-200). b , RMSD plot during MD simulations for the protein-protein complex.
Techniques Used:
Figure Legend Snippet: a , Illustration of the multiple-sequence alignment coverage of the OPTN-OPTN-PEX14 sequence used as input for predicting the complex structure. The black line represents the total coverage for each residue. Residues corresponding to OPTN 420–510 (proteins A and B) and PEX14 1-200 (protein C) are indicated. b , Plot of the pLDDT scores for the top-ranked 5 structural models. Each model is depicted in a different color. c , Free energy landscape, d , RMSF, e , hydrogen bond, f , gyrate, and g , SASA changes during molecular dynamics simulations.
Techniques Used: Sequencing, Residue
Figure Legend Snippet:
Techniques Used: Binding Assay, Protein-Protein interactions
Related Articles
Expressing:Article Title: PEX14 acts as the interface linking optineurin to cell type-selective pexophagy Article Snippet: .. The flippase (Flp) recombinase expression vector (pOG44; Thermo Fisher Scientific, V600520), the Flp-InTM inducible expression vector (pcDNA5/FRT/TO; Thermo Fisher Scientific, V652020), the lentiviral GFP expression vector (pLenti CMV GFP Puro; Addgene, plasmid 17448), and the plasmids encoding GFP (Clontech, 6085-1), the pOPINE GFP nanobody (Addgene, plasmid 49172 [ ]), Plasmid Preparation:Article Title: PEX14 acts as the interface linking optineurin to cell type-selective pexophagy Article Snippet: .. The flippase (Flp) recombinase expression vector (pOG44; Thermo Fisher Scientific, V600520), the Flp-InTM inducible expression vector (pcDNA5/FRT/TO; Thermo Fisher Scientific, V652020), the lentiviral GFP expression vector (pLenti CMV GFP Puro; Addgene, plasmid 17448), and the plasmids encoding GFP (Clontech, 6085-1), the pOPINE GFP nanobody (Addgene, plasmid 49172 [ ]), |