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pet 15b lfn dta  (Addgene inc)


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    Structured Review

    Addgene inc pet 15b lfn dta
    Pet 15b Lfn Dta, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pet+15b+lfn+dta/pET-15b+LFN-DTA+(Plasmid+%2311075)/pmc12053335-51-1-9
    Average 92 stars, based on 10 article reviews
    pet 15b lfn dta - by Bioz Stars, 2026-09
    92/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Antiviral siRNA delivered using attenuated, anthrax toxin protects cells from the cytopathic effects of Zika virus
    Article Snippet: .. The pET-15b LFn-DTA was a gift from John Collier (Addgene plasmid # 11,075). ..

    Article Title: A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
    Article Snippet: .. His 6 -LFn-MxiH WT and His 6 -LFn-MxiH 2A (containing point mutations V58A and I78A) were expressed in E. coli BL21(DE3) using vectors derived from pET-15b LFN-DTA, a gift from J. Collier (Harvard Medical School, Boston, MA)(Addgene plasmid #11075; ), and purified by Ni-NTA affinity purification and size-exclusion chromatography with a HiLoad 16/600 Superdex 75 pg column. .. Endotoxins were removed from PA and LFn-MxiH preparations using two extractions with Triton X-114, followed by removal of remaining detergent with Bio-Beads SM-2 beads (Bio-Rad Laboratories; ).

    Article Title: CRISPA: A Non-viral, Transient Cas9 Delivery System Based on Reengineered Anthrax Toxin
    Article Snippet: The Addgene plasmid #62933 (pET-Cas9-NLS-6xHis ( )) served as backbone for insertion of an LF N -encoding fragment upstream of the Cas9 sequence by seamless assembly (NEBuilder HiFi DNA Assembly Cloning Kit; #E5520; New England BioLabs GmbH, Frankfurt am Main, Germany). .. The LF N -encoding fragment was generated by PCR and with Addgene plasmid #11075 (pET-15b LFN-DTA ( )) as template. .. The NEBuilder Assembly Tool (New England BioLabs GmbH, Frankfurt am Main, Germany) was used to design primers for the seamless cloning procedure.

    Article Title: Antiviral siRNA delivered using attenuated, anthrax toxin protects cells from the cytopathic effects of Zika virus.
    Article Snippet: .. The pET-15b LFn-DTA was a gift from John Collier (Addgene plasmid # 11,075). ..

    Article Title: Antiviral siRNA Delivered Using Attenuated, Anthrax Toxin Protects Cells from the Cytopathic Effects of Zika Virus
    Article Snippet: .. The pET-15b LFn-DTA was a gift from John Collier (Addgene plasmid # 11075). ..

    Article Title: Method for preparing liposomes
    Article Snippet: .. The inventors loaded exosomes with the conditionally lethal cargo LFn-Diphtheria toxin A chain (DTA) using PA83 as described above, with the exception of the plasmid used, i.e. the plasmid was from Addgene (pET-15b LFn-DTA, Addgene number 11075) (https://www.addgene.org/11075/) Loading Exosomes The exosomes were loaded by incubating LFnDTA—SEQ ID No: 15 (protein) and SEQ ID No: 16 (DNA)—with HeLa cells at a concentration of 10 μg/ml and 50 μg/ml of SEQ ID No. 2 (PA83) for 4 hours at 37° C. in humidified atmosphere containing 5% (v/v) CO2. .. After this time the cells were washed with PBS and incubated with 5 μM Ionomycin (Sigma chemical company catalogue number I9657-1MG) in serum free media.

    Derivative Assay:

    Article Title: A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
    Article Snippet: .. His 6 -LFn-MxiH WT and His 6 -LFn-MxiH 2A (containing point mutations V58A and I78A) were expressed in E. coli BL21(DE3) using vectors derived from pET-15b LFN-DTA, a gift from J. Collier (Harvard Medical School, Boston, MA)(Addgene plasmid #11075; ), and purified by Ni-NTA affinity purification and size-exclusion chromatography with a HiLoad 16/600 Superdex 75 pg column. .. Endotoxins were removed from PA and LFn-MxiH preparations using two extractions with Triton X-114, followed by removal of remaining detergent with Bio-Beads SM-2 beads (Bio-Rad Laboratories; ).

    Purification:

    Article Title: A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
    Article Snippet: .. His 6 -LFn-MxiH WT and His 6 -LFn-MxiH 2A (containing point mutations V58A and I78A) were expressed in E. coli BL21(DE3) using vectors derived from pET-15b LFN-DTA, a gift from J. Collier (Harvard Medical School, Boston, MA)(Addgene plasmid #11075; ), and purified by Ni-NTA affinity purification and size-exclusion chromatography with a HiLoad 16/600 Superdex 75 pg column. .. Endotoxins were removed from PA and LFn-MxiH preparations using two extractions with Triton X-114, followed by removal of remaining detergent with Bio-Beads SM-2 beads (Bio-Rad Laboratories; ).

    Affinity Purification:

    Article Title: A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
    Article Snippet: .. His 6 -LFn-MxiH WT and His 6 -LFn-MxiH 2A (containing point mutations V58A and I78A) were expressed in E. coli BL21(DE3) using vectors derived from pET-15b LFN-DTA, a gift from J. Collier (Harvard Medical School, Boston, MA)(Addgene plasmid #11075; ), and purified by Ni-NTA affinity purification and size-exclusion chromatography with a HiLoad 16/600 Superdex 75 pg column. .. Endotoxins were removed from PA and LFn-MxiH preparations using two extractions with Triton X-114, followed by removal of remaining detergent with Bio-Beads SM-2 beads (Bio-Rad Laboratories; ).

    Size-exclusion Chromatography:

    Article Title: A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
    Article Snippet: .. His 6 -LFn-MxiH WT and His 6 -LFn-MxiH 2A (containing point mutations V58A and I78A) were expressed in E. coli BL21(DE3) using vectors derived from pET-15b LFN-DTA, a gift from J. Collier (Harvard Medical School, Boston, MA)(Addgene plasmid #11075; ), and purified by Ni-NTA affinity purification and size-exclusion chromatography with a HiLoad 16/600 Superdex 75 pg column. .. Endotoxins were removed from PA and LFn-MxiH preparations using two extractions with Triton X-114, followed by removal of remaining detergent with Bio-Beads SM-2 beads (Bio-Rad Laboratories; ).

    Generated:

    Article Title: CRISPA: A Non-viral, Transient Cas9 Delivery System Based on Reengineered Anthrax Toxin
    Article Snippet: The Addgene plasmid #62933 (pET-Cas9-NLS-6xHis ( )) served as backbone for insertion of an LF N -encoding fragment upstream of the Cas9 sequence by seamless assembly (NEBuilder HiFi DNA Assembly Cloning Kit; #E5520; New England BioLabs GmbH, Frankfurt am Main, Germany). .. The LF N -encoding fragment was generated by PCR and with Addgene plasmid #11075 (pET-15b LFN-DTA ( )) as template. .. The NEBuilder Assembly Tool (New England BioLabs GmbH, Frankfurt am Main, Germany) was used to design primers for the seamless cloning procedure.

    Polymerase Chain Reaction:

    Article Title: CRISPA: A Non-viral, Transient Cas9 Delivery System Based on Reengineered Anthrax Toxin
    Article Snippet: The Addgene plasmid #62933 (pET-Cas9-NLS-6xHis ( )) served as backbone for insertion of an LF N -encoding fragment upstream of the Cas9 sequence by seamless assembly (NEBuilder HiFi DNA Assembly Cloning Kit; #E5520; New England BioLabs GmbH, Frankfurt am Main, Germany). .. The LF N -encoding fragment was generated by PCR and with Addgene plasmid #11075 (pET-15b LFN-DTA ( )) as template. .. The NEBuilder Assembly Tool (New England BioLabs GmbH, Frankfurt am Main, Germany) was used to design primers for the seamless cloning procedure.

    Concentration Assay:

    Article Title: Method for preparing liposomes
    Article Snippet: .. The inventors loaded exosomes with the conditionally lethal cargo LFn-Diphtheria toxin A chain (DTA) using PA83 as described above, with the exception of the plasmid used, i.e. the plasmid was from Addgene (pET-15b LFn-DTA, Addgene number 11075) (https://www.addgene.org/11075/) Loading Exosomes The exosomes were loaded by incubating LFnDTA—SEQ ID No: 15 (protein) and SEQ ID No: 16 (DNA)—with HeLa cells at a concentration of 10 μg/ml and 50 μg/ml of SEQ ID No. 2 (PA83) for 4 hours at 37° C. in humidified atmosphere containing 5% (v/v) CO2. .. After this time the cells were washed with PBS and incubated with 5 μM Ionomycin (Sigma chemical company catalogue number I9657-1MG) in serum free media.



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