Western Blot:Article Title: MyoD1 localization at the nuclear periphery is mediated by association of WFS1 with active enhancers
Article Snippet: .. Primary antibodies used for immunoblotting and immunofluorescence experiments Antigen Host WB Dilution IF Dilution Source Lamin A/C Mouse 1:1000 1:100 Santa Cruz (clone E1) sc-376248 LAP2alpha Rabbit N/A 1:1000 245-23 Emerin Rabbit 1:1000 1:100 Atlas Antibodies (HPA000609) Lot #A114889 LBR Guinea pig 1:100 1:800 Gifted by Harald Herrmann HA Tag Mouse 1:1000 1:100 Sigma Aldrich (clone HA-7) H9658 FLAG Tag Mouse 1:2000 1:100 Sigma Aldrich (clone M2) F1804 γ-tubulin Mouse 1:5000 N/A Sigma Aldrich #T5326 (clone GTU- 88) WFS1 Rabbit 1:400 1:100 Proteintech (26995-1-AP) Cas9 Mouse 1:300 1:50 Generated by Stefan Schüchner, Max Perutz Labs, clone 7A9 RFP Rat N/A 1:1000 Proteintech (clone 5F8) RFP Mouse 1:2000 N/A Proteintech (clone 6G6) LEMD2 Mouse 1:2500 1:100 Atlas Antibodies (HPA017340) Lot #A60701 LAP2beta Rabbit 1:10 Undiluted Generated by C. Harris (mAK17, aa313-330) H3K9me3 Rabbit 1:1000 1:800 Abcam (ab8898) Lot #GR3365652-1 H3 Rabbit 1:1000 N/A Abcam (ab1791) Lot #GR153323-6 H3K9me2 Rabbit 1:1000 1:800 Active Motif (39420) Lot #34718002 H3K27me3 Rabbit 1:1000 N/A Merck-Millipore (07-449) Lot #2275589 MyoD1 Mouse 1:600 N/A Santa Cruz (clone G-1) sc-77460 Myogenin Mouse 1:50 1:10 Santa Cruz (clone F5D) sc-12732 MyHC Mouse 1:50 1:10 Invitrogen (clone MF20) 14-6503-82 Lamin B1 Rabbit 1:500 1:50 Proteintech (12987-1-AP) Lot #44- 161-2324577 Supplementary Table 8. .. PCRs for vector assembly Assembly 1 Fragment Forward primer Reverse primer Template PGK promoter PGK promoter-F PGK promoter-R Kindly gifted from Nicolaus Beer FLAG Tag FLAG Tag-F FLAG Tag-R Kindly gifted from Daria Filipczak WFS1 coding sequence A WFS1 CDS A-F WFS1 CDS A-R Twist Bioscience sequence A WFS1 coding sequence B WFS1 CDS B-F WFS1 CDS B-R Twist Bioscience sequence B Assembly 2 Fragment Forward primer Reverse primer Template PGK promoter PGK promoter-F PGK promoter-R Kindly gifted from Nicolaus Beer FLAG Tag FLAG Tag-F FLAG Tag-R2 Kindly gifted from Daria Filipczak WFS1 coding sequence WFS1 CDS HA-F WFS1 CDS HA-R WFS1-FLAG Tag plasmid mScarlet mScarlet-F mScarlet-R Addgene #85042 The HA Tag for assembly 2 was added as a flanking sequence to the primers used for PCR amplification.
Immunofluorescence:Article Title: MyoD1 localization at the nuclear periphery is mediated by association of WFS1 with active enhancers
Article Snippet: .. Primary antibodies used for immunoblotting and immunofluorescence experiments Antigen Host WB Dilution IF Dilution Source Lamin A/C Mouse 1:1000 1:100 Santa Cruz (clone E1) sc-376248 LAP2alpha Rabbit N/A 1:1000 245-23 Emerin Rabbit 1:1000 1:100 Atlas Antibodies (HPA000609) Lot #A114889 LBR Guinea pig 1:100 1:800 Gifted by Harald Herrmann HA Tag Mouse 1:1000 1:100 Sigma Aldrich (clone HA-7) H9658 FLAG Tag Mouse 1:2000 1:100 Sigma Aldrich (clone M2) F1804 γ-tubulin Mouse 1:5000 N/A Sigma Aldrich #T5326 (clone GTU- 88) WFS1 Rabbit 1:400 1:100 Proteintech (26995-1-AP) Cas9 Mouse 1:300 1:50 Generated by Stefan Schüchner, Max Perutz Labs, clone 7A9 RFP Rat N/A 1:1000 Proteintech (clone 5F8) RFP Mouse 1:2000 N/A Proteintech (clone 6G6) LEMD2 Mouse 1:2500 1:100 Atlas Antibodies (HPA017340) Lot #A60701 LAP2beta Rabbit 1:10 Undiluted Generated by C. Harris (mAK17, aa313-330) H3K9me3 Rabbit 1:1000 1:800 Abcam (ab8898) Lot #GR3365652-1 H3 Rabbit 1:1000 N/A Abcam (ab1791) Lot #GR153323-6 H3K9me2 Rabbit 1:1000 1:800 Active Motif (39420) Lot #34718002 H3K27me3 Rabbit 1:1000 N/A Merck-Millipore (07-449) Lot #2275589 MyoD1 Mouse 1:600 N/A Santa Cruz (clone G-1) sc-77460 Myogenin Mouse 1:50 1:10 Santa Cruz (clone F5D) sc-12732 MyHC Mouse 1:50 1:10 Invitrogen (clone MF20) 14-6503-82 Lamin B1 Rabbit 1:500 1:50 Proteintech (12987-1-AP) Lot #44- 161-2324577 Supplementary Table 8. .. PCRs for vector assembly Assembly 1 Fragment Forward primer Reverse primer Template PGK promoter PGK promoter-F PGK promoter-R Kindly gifted from Nicolaus Beer FLAG Tag FLAG Tag-F FLAG Tag-R Kindly gifted from Daria Filipczak WFS1 coding sequence A WFS1 CDS A-F WFS1 CDS A-R Twist Bioscience sequence A WFS1 coding sequence B WFS1 CDS B-F WFS1 CDS B-R Twist Bioscience sequence B Assembly 2 Fragment Forward primer Reverse primer Template PGK promoter PGK promoter-F PGK promoter-R Kindly gifted from Nicolaus Beer FLAG Tag FLAG Tag-F FLAG Tag-R2 Kindly gifted from Daria Filipczak WFS1 coding sequence WFS1 CDS HA-F WFS1 CDS HA-R WFS1-FLAG Tag plasmid mScarlet mScarlet-F mScarlet-R Addgene #85042 The HA Tag for assembly 2 was added as a flanking sequence to the primers used for PCR amplification.
FLAG-tag:Article Title: MyoD1 localization at the nuclear periphery is mediated by association of WFS1 with active enhancers
Article Snippet: .. Primary antibodies used for immunoblotting and immunofluorescence experiments Antigen Host WB Dilution IF Dilution Source Lamin A/C Mouse 1:1000 1:100 Santa Cruz (clone E1) sc-376248 LAP2alpha Rabbit N/A 1:1000 245-23 Emerin Rabbit 1:1000 1:100 Atlas Antibodies (HPA000609) Lot #A114889 LBR Guinea pig 1:100 1:800 Gifted by Harald Herrmann HA Tag Mouse 1:1000 1:100 Sigma Aldrich (clone HA-7) H9658 FLAG Tag Mouse 1:2000 1:100 Sigma Aldrich (clone M2) F1804 γ-tubulin Mouse 1:5000 N/A Sigma Aldrich #T5326 (clone GTU- 88) WFS1 Rabbit 1:400 1:100 Proteintech (26995-1-AP) Cas9 Mouse 1:300 1:50 Generated by Stefan Schüchner, Max Perutz Labs, clone 7A9 RFP Rat N/A 1:1000 Proteintech (clone 5F8) RFP Mouse 1:2000 N/A Proteintech (clone 6G6) LEMD2 Mouse 1:2500 1:100 Atlas Antibodies (HPA017340) Lot #A60701 LAP2beta Rabbit 1:10 Undiluted Generated by C. Harris (mAK17, aa313-330) H3K9me3 Rabbit 1:1000 1:800 Abcam (ab8898) Lot #GR3365652-1 H3 Rabbit 1:1000 N/A Abcam (ab1791) Lot #GR153323-6 H3K9me2 Rabbit 1:1000 1:800 Active Motif (39420) Lot #34718002 H3K27me3 Rabbit 1:1000 N/A Merck-Millipore (07-449) Lot #2275589 MyoD1 Mouse 1:600 N/A Santa Cruz (clone G-1) sc-77460 Myogenin Mouse 1:50 1:10 Santa Cruz (clone F5D) sc-12732 MyHC Mouse 1:50 1:10 Invitrogen (clone MF20) 14-6503-82 Lamin B1 Rabbit 1:500 1:50 Proteintech (12987-1-AP) Lot #44- 161-2324577 Supplementary Table 8. .. PCRs for vector assembly Assembly 1 Fragment Forward primer Reverse primer Template PGK promoter PGK promoter-F PGK promoter-R Kindly gifted from Nicolaus Beer FLAG Tag FLAG Tag-F FLAG Tag-R Kindly gifted from Daria Filipczak WFS1 coding sequence A WFS1 CDS A-F WFS1 CDS A-R Twist Bioscience sequence A WFS1 coding sequence B WFS1 CDS B-F WFS1 CDS B-R Twist Bioscience sequence B Assembly 2 Fragment Forward primer Reverse primer Template PGK promoter PGK promoter-F PGK promoter-R Kindly gifted from Nicolaus Beer FLAG Tag FLAG Tag-F FLAG Tag-R2 Kindly gifted from Daria Filipczak WFS1 coding sequence WFS1 CDS HA-F WFS1 CDS HA-R WFS1-FLAG Tag plasmid mScarlet mScarlet-F mScarlet-R Addgene #85042 The HA Tag for assembly 2 was added as a flanking sequence to the primers used for PCR amplification.
Generated:Article Title: MyoD1 localization at the nuclear periphery is mediated by association of WFS1 with active enhancers
Article Snippet: .. Primary antibodies used for immunoblotting and immunofluorescence experiments Antigen Host WB Dilution IF Dilution Source Lamin A/C Mouse 1:1000 1:100 Santa Cruz (clone E1) sc-376248 LAP2alpha Rabbit N/A 1:1000 245-23 Emerin Rabbit 1:1000 1:100 Atlas Antibodies (HPA000609) Lot #A114889 LBR Guinea pig 1:100 1:800 Gifted by Harald Herrmann HA Tag Mouse 1:1000 1:100 Sigma Aldrich (clone HA-7) H9658 FLAG Tag Mouse 1:2000 1:100 Sigma Aldrich (clone M2) F1804 γ-tubulin Mouse 1:5000 N/A Sigma Aldrich #T5326 (clone GTU- 88) WFS1 Rabbit 1:400 1:100 Proteintech (26995-1-AP) Cas9 Mouse 1:300 1:50 Generated by Stefan Schüchner, Max Perutz Labs, clone 7A9 RFP Rat N/A 1:1000 Proteintech (clone 5F8) RFP Mouse 1:2000 N/A Proteintech (clone 6G6) LEMD2 Mouse 1:2500 1:100 Atlas Antibodies (HPA017340) Lot #A60701 LAP2beta Rabbit 1:10 Undiluted Generated by C. Harris (mAK17, aa313-330) H3K9me3 Rabbit 1:1000 1:800 Abcam (ab8898) Lot #GR3365652-1 H3 Rabbit 1:1000 N/A Abcam (ab1791) Lot #GR153323-6 H3K9me2 Rabbit 1:1000 1:800 Active Motif (39420) Lot #34718002 H3K27me3 Rabbit 1:1000 N/A Merck-Millipore (07-449) Lot #2275589 MyoD1 Mouse 1:600 N/A Santa Cruz (clone G-1) sc-77460 Myogenin Mouse 1:50 1:10 Santa Cruz (clone F5D) sc-12732 MyHC Mouse 1:50 1:10 Invitrogen (clone MF20) 14-6503-82 Lamin B1 Rabbit 1:500 1:50 Proteintech (12987-1-AP) Lot #44- 161-2324577 Supplementary Table 8. .. PCRs for vector assembly Assembly 1 Fragment Forward primer Reverse primer Template PGK promoter PGK promoter-F PGK promoter-R Kindly gifted from Nicolaus Beer FLAG Tag FLAG Tag-F FLAG Tag-R Kindly gifted from Daria Filipczak WFS1 coding sequence A WFS1 CDS A-F WFS1 CDS A-R Twist Bioscience sequence A WFS1 coding sequence B WFS1 CDS B-F WFS1 CDS B-R Twist Bioscience sequence B Assembly 2 Fragment Forward primer Reverse primer Template PGK promoter PGK promoter-F PGK promoter-R Kindly gifted from Nicolaus Beer FLAG Tag FLAG Tag-F FLAG Tag-R2 Kindly gifted from Daria Filipczak WFS1 coding sequence WFS1 CDS HA-F WFS1 CDS HA-R WFS1-FLAG Tag plasmid mScarlet mScarlet-F mScarlet-R Addgene #85042 The HA Tag for assembly 2 was added as a flanking sequence to the primers used for PCR amplification.
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