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flag  (Addgene inc)


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    Structured Review

    Addgene inc flag
    Flag, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pyd+flag/pcDNA3-N-Flag-ASC1+(Plasmid+%2375134)/pm38486019-331-22-27
    Average 93 stars, based on 14 article reviews
    flag - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Over Expression:

    Article Title: Anaphylactic degranulation by mast cells requires the mobilization of inflammasome components.
    Article Snippet: The images were captured using a GelDoc touch screen (Bio-Rad). .. For analyzing overexpression protein interaction studies, GFP-CD63 (10 μg; Sino Biological, MG50557-ANG), Flag–NLRP3/Flag– PYD/Flag–NBD/Flag–LRR (10 μg each; Addgene, 75137, 75140, 75141), and Flag–ASC/Flag–PYD/Flag–CARD truncate (10 μg each; Addgene, 75134); or GFP-CD63 full length/or its truncates, GFP-tagged TM1, TM2, TM3 and TM4, or 1 and 5 μg of Flag-tagged NLRP3 or Myc-tagged ASC with a constant amount of GFP-CD63-TM3 (5 μg) were transfected into HEK293T cells (700,000 cells per well) using JetPrime transfection reagent (Polyplus). .. After 36 h, cell lysates were lysed and incubated with 1 μg of Flag (Cell Signaling, 8146) or GFP (Santa Cruz, sc-9996) or Myc (Cell Signaling, 5605) antibodies.

    Transfection:

    Article Title: Anaphylactic degranulation by mast cells requires the mobilization of inflammasome components.
    Article Snippet: The images were captured using a GelDoc touch screen (Bio-Rad). .. For analyzing overexpression protein interaction studies, GFP-CD63 (10 μg; Sino Biological, MG50557-ANG), Flag–NLRP3/Flag– PYD/Flag–NBD/Flag–LRR (10 μg each; Addgene, 75137, 75140, 75141), and Flag–ASC/Flag–PYD/Flag–CARD truncate (10 μg each; Addgene, 75134); or GFP-CD63 full length/or its truncates, GFP-tagged TM1, TM2, TM3 and TM4, or 1 and 5 μg of Flag-tagged NLRP3 or Myc-tagged ASC with a constant amount of GFP-CD63-TM3 (5 μg) were transfected into HEK293T cells (700,000 cells per well) using JetPrime transfection reagent (Polyplus). .. After 36 h, cell lysates were lysed and incubated with 1 μg of Flag (Cell Signaling, 8146) or GFP (Santa Cruz, sc-9996) or Myc (Cell Signaling, 5605) antibodies.



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    High-throughput screening of a 9,600 core chemical library using NLRP3 inflammasome in a cell-free system. ( a ) Schematic representations of C-terminal biotinylated full-length NLRP3 (NLRP3-FL-Btn), the N-terminal <t>FLAG-tagged</t> PYD <t>of</t> <t>ASC</t> (FLAG-ASC-PYD), and the N-terminal FLAG-tagged caspase recruit domain (CARD) of ASC (FLAG-ASC-CARD). PYD pyrin domain, CARD caspase recruitment domain, NOD nucleotide-binding oligomerization domain, LRR leucine rich repeat. ( b ) Schematic representation of reconstituted inflammasomes. The PYD of truncated ASC was able to open and bind to the PYD of NLRP3. The chemical energy of the reactive oxygen on donor beads was transferred to acceptor beads, and a signal was detected. ( c ) Primary screening of the 9,600 core chemical library using NLRP3 inflammasome in a cell-free system. The result presented was the only result obtained.
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    High-throughput screening of a 9,600 core chemical library using NLRP3 inflammasome in a cell-free system. ( a ) Schematic representations of C-terminal biotinylated full-length NLRP3 (NLRP3-FL-Btn), the N-terminal <t>FLAG-tagged</t> PYD <t>of</t> <t>ASC</t> (FLAG-ASC-PYD), and the N-terminal FLAG-tagged caspase recruit domain (CARD) of ASC (FLAG-ASC-CARD). PYD pyrin domain, CARD caspase recruitment domain, NOD nucleotide-binding oligomerization domain, LRR leucine rich repeat. ( b ) Schematic representation of reconstituted inflammasomes. The PYD of truncated ASC was able to open and bind to the PYD of NLRP3. The chemical energy of the reactive oxygen on donor beads was transferred to acceptor beads, and a signal was detected. ( c ) Primary screening of the 9,600 core chemical library using NLRP3 inflammasome in a cell-free system. The result presented was the only result obtained.
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    Image Search Results


    High-throughput screening of a 9,600 core chemical library using NLRP3 inflammasome in a cell-free system. ( a ) Schematic representations of C-terminal biotinylated full-length NLRP3 (NLRP3-FL-Btn), the N-terminal FLAG-tagged PYD of ASC (FLAG-ASC-PYD), and the N-terminal FLAG-tagged caspase recruit domain (CARD) of ASC (FLAG-ASC-CARD). PYD pyrin domain, CARD caspase recruitment domain, NOD nucleotide-binding oligomerization domain, LRR leucine rich repeat. ( b ) Schematic representation of reconstituted inflammasomes. The PYD of truncated ASC was able to open and bind to the PYD of NLRP3. The chemical energy of the reactive oxygen on donor beads was transferred to acceptor beads, and a signal was detected. ( c ) Primary screening of the 9,600 core chemical library using NLRP3 inflammasome in a cell-free system. The result presented was the only result obtained.

    Journal: Scientific Reports

    Article Title: KN3014, a piperidine-containing small compound, inhibits auto-secretion of IL-1β from PBMCs in a patient with Muckle–Wells syndrome

    doi: 10.1038/s41598-020-70513-0

    Figure Lengend Snippet: High-throughput screening of a 9,600 core chemical library using NLRP3 inflammasome in a cell-free system. ( a ) Schematic representations of C-terminal biotinylated full-length NLRP3 (NLRP3-FL-Btn), the N-terminal FLAG-tagged PYD of ASC (FLAG-ASC-PYD), and the N-terminal FLAG-tagged caspase recruit domain (CARD) of ASC (FLAG-ASC-CARD). PYD pyrin domain, CARD caspase recruitment domain, NOD nucleotide-binding oligomerization domain, LRR leucine rich repeat. ( b ) Schematic representation of reconstituted inflammasomes. The PYD of truncated ASC was able to open and bind to the PYD of NLRP3. The chemical energy of the reactive oxygen on donor beads was transferred to acceptor beads, and a signal was detected. ( c ) Primary screening of the 9,600 core chemical library using NLRP3 inflammasome in a cell-free system. The result presented was the only result obtained.

    Article Snippet: In addition, 10 µL of an ASC-PYD mixture containing 1 µL FLAG-ASC-PYD was added to sample wells and positive control wells using a FlexDrop dispenser, and 10 µL of an ASC-CARD mixture containing 1 µL FLAG-ASC-CARD was added to negative control wells using a Picus NxT dispenser (Sartorius, Goettingen, Germany).

    Techniques: High Throughput Screening Assay, Binding Assay