Immunofluorescence:Article Title: A rapid inducible RNA decay system reveals fast mRNA decay in P-bodies
Article Snippet: .. For smFISH combined with immunofluorescence, 1:1000 dilution of rabbit anti-DCP1a (Abcam ab183709), 1:100 rabbit anti-XRN1 (Bethyl Laboratories A300-443A-M), 1:1000 rabbit anti-DDX6 (Bethyl Laboratories A300-461A), or 1:100 rabbit anti-G3BP (Aviva Systems Biology ARP37713_T100) were used as primary antibodies. .. A 1:5000 dilution of goat-anti-rabbit IgG (H + L) Alexa Fluor 750, Invitrogen A-21039, was used as the secondary antibody for all primary rabbit-derived antibodies.
Article Title: p62 sorts Lupus La and selected microRNAs into breast cancer-derived exosomes
Article Snippet: Secondary antibodies used for immunoblot (1:10,000 dilution) were HRP-linked sheep anti-mouse IgG (NA931; Cytiva) and HRP-linked donkey anti-rabbit IgG (NA934; Cytiva). .. Primary antibodies used for immunofluorescence (1:100 dilution) were mouse anti-La/SSB (OTI1E11; OriGene), rabbit anti-La/SSB (D19B3; Cell Signaling Technology), mouse anti-CD63 (556019; BD Biosciences), rabbit anti-DDX6 (A300-461A; Bethyl Laboratories), rabbit anti-LC3B (NB100-2220; Novus Biologicals), and guinea pig anti-p62/SQSTM1 (GP62-C; Biogen Biotechnik). .. Primary antibodies used for immunoblot (1:1,000 dilution) were mouse anti-ALIX (sc-53540; Santa Cruz), rabbit anti-annexin A2 (ab185957; Abcam), rabbit anti-ATG7 (D12B11; Cell Signaling Technology), mouse anti-CD63 (556019; BD Biosciences), rabbit anti-FIP200 (aka RB1CC1, 17250-1-AP; Proteintech), mouse anti-flotillin 2 (610384; BD Biosciences), rabbit anti-HA-Tag (C29F4; Cell Signaling Technology), mouse anti-La/SSB (OTI1E11; OriGene), rabbit anti-La/SSB (D19B3; Cell Signaling Technology), rabbit anti-LAMP1 (D2D11; Cell Signaling Technology), rabbit anti-LAMP2A (ab18528; Abcam), mouse anti-LC3B (E5Q2K; Cell Signaling Technology), mouse anti-p62/SQSTM1 (ab56416; Abcam), rabbit anti-p62/SQSTM1 (PM045; MBL International), rabbit anti-Rab5 (D4F5; Cell Signaling Technology), mouse anti-TIM23 (611222; BD Biosciences), and rabbit anti-vinculin (ab129002; Abcam).
Article Title: A rapid inducible RNA decay system reveals fast mRNA decay in P-bodies.
Article Snippet: .. For smFISH combined with immunofluorescence, 1:1000 dilution of rabbit anti-DCP1a (Abcam ab183709), 1:100 rabbit anti-XRN1 (Bethyl Laboratories A300-443A-M), 1:1000 rabbit anti-DDX6 (Bethyl Laboratories A300-461A), or 1:100 rabbit anti-G3BP (Aviva Systems Biology ARP37713_T100) were used as primary antibodies. .. A 1:5000 dilution of goat-anti-rabbit IgG (H + L) Alexa Fluor 750, Invitrogen A21039, was used as the secondary antibody for all primary rabbitderived antibodies.
Article Title: p62 sorts Lupus La and selected microRNAs into breast cancer-derived exosomes
Article Snippet: Secondary antibodies used for immunoblot (1:10,000 dilution) were: HRP-linked sheep anti-mouse IgG (Cytiva NA931) and HRP-linked donkey anti-rabbit IgG (Cytiva NA934). .. Primary antibodies used for immunofluorescence (1:100 dilution) were: mouse anti-La/SSB (OriGene OTI1E11), rabbit anti-La/SSB (Cell Signaling Technology D19B3), mouse anti-CD63 (BD Biosciences 556019), rabbit anti-DDX6 (Bethyl Laboratories A300-461A) and guinea pig anti-p62/SQSTM1 (Biogen Biotechnik GP62-C). .. Secondary antibodies used for immunofluorescence (1:500 dilution) were: AlexaFluor 488 donkey anti-rabbit IgG (Invitrogen A-21202), AlexaFluor 555 goat anti-mouse IgG (Invitrogen A-21424) and AlexaFluor 647 goat anti-guinea pig IgG (Invitrogen A-21450).
Hemagglutination Assay:Article Title: DDX6 interacts with DDX3X to repress translation in microRNA-mediated silencing
Article Snippet: .. CXCR miRNA: TOP: 5’-GUU UUC ACA AAG CUA ACA CA-3’ CXCR miRNA BOTTOM: 5’-UGU UAG CUG GAG UGA AAA CUU-3’ mir-30 TOP: GCU GUA AAC AUC CUC GAC UGG AAG UU mir-30 BOTTOM: CUU UCA GUC GGA UGU UUG CAG CUU Table S3: Antibodies used in this paper DDX6 Bethyl A310-131A, rabbit anti-DDX6, 1:5000, MW 55 KDa Bethyl A310-460A, rabbit anti-DDX6, 1:5000 DDX3X Bethyl A300-474A, rabbit anti-DDX3, 1:5000, MW 75kDa Flag Sigma F3165, Anti-Flag M2 Monoclonal Antibody, 1:2000 HA Sigma H6908, rabbit anti-HA antibody, 1:2000 Roche Appplied Science, 11 583 816 001, Mouse, 1:2000 V5 AbD Serotec Clone SV5-PK1 mouse, 1:2000 Myc DSHB, 9E10 mouse antibody 1:20 GFP Invitrogen, A-11122, rabbit antibody, 1:1000 Bim Enzo Life Sciences, ADI-AAP-330, rabbit anti-Bim/BOD polyclonal Ab, 1:1000, MW 23, 16, 13 kDa Bim Cell Signaling 2933, rabbit mAb (C34C5), 1;1000, MW 23, 16, 13 kDa 4E-T Fortis, A300-706A, rabbit anti-4E-T/eIF4E-T antibody affinity purified, 1:1000, 110 KDa His Novagen 70796-4, His Tag Monoclonal antibody, 1:4000 GST Proteintech, 66001-1-lg, Mouse antibody, 1:50000, MW 26kDa Caspase 2 Cell Signaling #2224, mouse antibody, 1:1000, MW 12, 14, 48kDa GAPDH GAPDH (6C5) antibody Santa Cruz Biotechnology Cat# sc-32233, RRID:AB_627679 Dcp2 Bethyl A302-597A, Rabbit anti-Dcp2, 1:2000, MW about 55kDa Puromycin Mouse Anti-Puromycin antibody (3RH11), KeraFAST, 1:2000 eIF3A Cell Signaling 3411, anti-eIF3A (D51F4) XP TM Rabbit mAb, 1:1000, MW 166 kDa RPL7 Bethyl, A300-740A, MW 29kDa Tubulin Beta III isoform Millipore MAB1637, Mouse, 1:2000, MW 55 kDa Flag Anti-FLAG M2 Affinity Gel, Sigma, A2220 Total protein quantification LI-COR P/N 926-11010, Revert 700 Total protein Stain Kit HRP-conjugated secondary antibody Goat anti-Mouse IgG(H/L):HRP, BioRad 5178-2504, 1:100-1:5000 Goat anti-rabbit IgG(H/L):HRP, BioRad, 5196-2504 , 1:1000-1:5000 DyLight 800 secondary antibody Invitrogen SA5-35571, goat anti-Rabbit IgG DyLight 800 FITC- or Cy3 conjugated secondary antibody CyTM3-conjugated AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch, 711-165-152, 1:200 Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG (H+L), Jackson ImmunoResearch 115-545-166, 1:200 Alexa Fluor 488-conjugated AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch 711-545-152, 1:200 Figure S1. ..
Cell-Signaling:Article Title: DDX6 interacts with DDX3X to repress translation in microRNA-mediated silencing
Article Snippet: .. CXCR miRNA: TOP: 5’-GUU UUC ACA AAG CUA ACA CA-3’ CXCR miRNA BOTTOM: 5’-UGU UAG CUG GAG UGA AAA CUU-3’ mir-30 TOP: GCU GUA AAC AUC CUC GAC UGG AAG UU mir-30 BOTTOM: CUU UCA GUC GGA UGU UUG CAG CUU Table S3: Antibodies used in this paper DDX6 Bethyl A310-131A, rabbit anti-DDX6, 1:5000, MW 55 KDa Bethyl A310-460A, rabbit anti-DDX6, 1:5000 DDX3X Bethyl A300-474A, rabbit anti-DDX3, 1:5000, MW 75kDa Flag Sigma F3165, Anti-Flag M2 Monoclonal Antibody, 1:2000 HA Sigma H6908, rabbit anti-HA antibody, 1:2000 Roche Appplied Science, 11 583 816 001, Mouse, 1:2000 V5 AbD Serotec Clone SV5-PK1 mouse, 1:2000 Myc DSHB, 9E10 mouse antibody 1:20 GFP Invitrogen, A-11122, rabbit antibody, 1:1000 Bim Enzo Life Sciences, ADI-AAP-330, rabbit anti-Bim/BOD polyclonal Ab, 1:1000, MW 23, 16, 13 kDa Bim Cell Signaling 2933, rabbit mAb (C34C5), 1;1000, MW 23, 16, 13 kDa 4E-T Fortis, A300-706A, rabbit anti-4E-T/eIF4E-T antibody affinity purified, 1:1000, 110 KDa His Novagen 70796-4, His Tag Monoclonal antibody, 1:4000 GST Proteintech, 66001-1-lg, Mouse antibody, 1:50000, MW 26kDa Caspase 2 Cell Signaling #2224, mouse antibody, 1:1000, MW 12, 14, 48kDa GAPDH GAPDH (6C5) antibody Santa Cruz Biotechnology Cat# sc-32233, RRID:AB_627679 Dcp2 Bethyl A302-597A, Rabbit anti-Dcp2, 1:2000, MW about 55kDa Puromycin Mouse Anti-Puromycin antibody (3RH11), KeraFAST, 1:2000 eIF3A Cell Signaling 3411, anti-eIF3A (D51F4) XP TM Rabbit mAb, 1:1000, MW 166 kDa RPL7 Bethyl, A300-740A, MW 29kDa Tubulin Beta III isoform Millipore MAB1637, Mouse, 1:2000, MW 55 kDa Flag Anti-FLAG M2 Affinity Gel, Sigma, A2220 Total protein quantification LI-COR P/N 926-11010, Revert 700 Total protein Stain Kit HRP-conjugated secondary antibody Goat anti-Mouse IgG(H/L):HRP, BioRad 5178-2504, 1:100-1:5000 Goat anti-rabbit IgG(H/L):HRP, BioRad, 5196-2504 , 1:1000-1:5000 DyLight 800 secondary antibody Invitrogen SA5-35571, goat anti-Rabbit IgG DyLight 800 FITC- or Cy3 conjugated secondary antibody CyTM3-conjugated AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch, 711-165-152, 1:200 Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG (H+L), Jackson ImmunoResearch 115-545-166, 1:200 Alexa Fluor 488-conjugated AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch 711-545-152, 1:200 Figure S1. ..
Affinity Purification:Article Title: DDX6 interacts with DDX3X to repress translation in microRNA-mediated silencing
Article Snippet: .. CXCR miRNA: TOP: 5’-GUU UUC ACA AAG CUA ACA CA-3’ CXCR miRNA BOTTOM: 5’-UGU UAG CUG GAG UGA AAA CUU-3’ mir-30 TOP: GCU GUA AAC AUC CUC GAC UGG AAG UU mir-30 BOTTOM: CUU UCA GUC GGA UGU UUG CAG CUU Table S3: Antibodies used in this paper DDX6 Bethyl A310-131A, rabbit anti-DDX6, 1:5000, MW 55 KDa Bethyl A310-460A, rabbit anti-DDX6, 1:5000 DDX3X Bethyl A300-474A, rabbit anti-DDX3, 1:5000, MW 75kDa Flag Sigma F3165, Anti-Flag M2 Monoclonal Antibody, 1:2000 HA Sigma H6908, rabbit anti-HA antibody, 1:2000 Roche Appplied Science, 11 583 816 001, Mouse, 1:2000 V5 AbD Serotec Clone SV5-PK1 mouse, 1:2000 Myc DSHB, 9E10 mouse antibody 1:20 GFP Invitrogen, A-11122, rabbit antibody, 1:1000 Bim Enzo Life Sciences, ADI-AAP-330, rabbit anti-Bim/BOD polyclonal Ab, 1:1000, MW 23, 16, 13 kDa Bim Cell Signaling 2933, rabbit mAb (C34C5), 1;1000, MW 23, 16, 13 kDa 4E-T Fortis, A300-706A, rabbit anti-4E-T/eIF4E-T antibody affinity purified, 1:1000, 110 KDa His Novagen 70796-4, His Tag Monoclonal antibody, 1:4000 GST Proteintech, 66001-1-lg, Mouse antibody, 1:50000, MW 26kDa Caspase 2 Cell Signaling #2224, mouse antibody, 1:1000, MW 12, 14, 48kDa GAPDH GAPDH (6C5) antibody Santa Cruz Biotechnology Cat# sc-32233, RRID:AB_627679 Dcp2 Bethyl A302-597A, Rabbit anti-Dcp2, 1:2000, MW about 55kDa Puromycin Mouse Anti-Puromycin antibody (3RH11), KeraFAST, 1:2000 eIF3A Cell Signaling 3411, anti-eIF3A (D51F4) XP TM Rabbit mAb, 1:1000, MW 166 kDa RPL7 Bethyl, A300-740A, MW 29kDa Tubulin Beta III isoform Millipore MAB1637, Mouse, 1:2000, MW 55 kDa Flag Anti-FLAG M2 Affinity Gel, Sigma, A2220 Total protein quantification LI-COR P/N 926-11010, Revert 700 Total protein Stain Kit HRP-conjugated secondary antibody Goat anti-Mouse IgG(H/L):HRP, BioRad 5178-2504, 1:100-1:5000 Goat anti-rabbit IgG(H/L):HRP, BioRad, 5196-2504 , 1:1000-1:5000 DyLight 800 secondary antibody Invitrogen SA5-35571, goat anti-Rabbit IgG DyLight 800 FITC- or Cy3 conjugated secondary antibody CyTM3-conjugated AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch, 711-165-152, 1:200 Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG (H+L), Jackson ImmunoResearch 115-545-166, 1:200 Alexa Fluor 488-conjugated AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch 711-545-152, 1:200 Figure S1. ..
Staining:Article Title: DDX6 interacts with DDX3X to repress translation in microRNA-mediated silencing
Article Snippet: .. CXCR miRNA: TOP: 5’-GUU UUC ACA AAG CUA ACA CA-3’ CXCR miRNA BOTTOM: 5’-UGU UAG CUG GAG UGA AAA CUU-3’ mir-30 TOP: GCU GUA AAC AUC CUC GAC UGG AAG UU mir-30 BOTTOM: CUU UCA GUC GGA UGU UUG CAG CUU Table S3: Antibodies used in this paper DDX6 Bethyl A310-131A, rabbit anti-DDX6, 1:5000, MW 55 KDa Bethyl A310-460A, rabbit anti-DDX6, 1:5000 DDX3X Bethyl A300-474A, rabbit anti-DDX3, 1:5000, MW 75kDa Flag Sigma F3165, Anti-Flag M2 Monoclonal Antibody, 1:2000 HA Sigma H6908, rabbit anti-HA antibody, 1:2000 Roche Appplied Science, 11 583 816 001, Mouse, 1:2000 V5 AbD Serotec Clone SV5-PK1 mouse, 1:2000 Myc DSHB, 9E10 mouse antibody 1:20 GFP Invitrogen, A-11122, rabbit antibody, 1:1000 Bim Enzo Life Sciences, ADI-AAP-330, rabbit anti-Bim/BOD polyclonal Ab, 1:1000, MW 23, 16, 13 kDa Bim Cell Signaling 2933, rabbit mAb (C34C5), 1;1000, MW 23, 16, 13 kDa 4E-T Fortis, A300-706A, rabbit anti-4E-T/eIF4E-T antibody affinity purified, 1:1000, 110 KDa His Novagen 70796-4, His Tag Monoclonal antibody, 1:4000 GST Proteintech, 66001-1-lg, Mouse antibody, 1:50000, MW 26kDa Caspase 2 Cell Signaling #2224, mouse antibody, 1:1000, MW 12, 14, 48kDa GAPDH GAPDH (6C5) antibody Santa Cruz Biotechnology Cat# sc-32233, RRID:AB_627679 Dcp2 Bethyl A302-597A, Rabbit anti-Dcp2, 1:2000, MW about 55kDa Puromycin Mouse Anti-Puromycin antibody (3RH11), KeraFAST, 1:2000 eIF3A Cell Signaling 3411, anti-eIF3A (D51F4) XP TM Rabbit mAb, 1:1000, MW 166 kDa RPL7 Bethyl, A300-740A, MW 29kDa Tubulin Beta III isoform Millipore MAB1637, Mouse, 1:2000, MW 55 kDa Flag Anti-FLAG M2 Affinity Gel, Sigma, A2220 Total protein quantification LI-COR P/N 926-11010, Revert 700 Total protein Stain Kit HRP-conjugated secondary antibody Goat anti-Mouse IgG(H/L):HRP, BioRad 5178-2504, 1:100-1:5000 Goat anti-rabbit IgG(H/L):HRP, BioRad, 5196-2504 , 1:1000-1:5000 DyLight 800 secondary antibody Invitrogen SA5-35571, goat anti-Rabbit IgG DyLight 800 FITC- or Cy3 conjugated secondary antibody CyTM3-conjugated AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch, 711-165-152, 1:200 Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG (H+L), Jackson ImmunoResearch 115-545-166, 1:200 Alexa Fluor 488-conjugated AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch 711-545-152, 1:200 Figure S1. ..
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