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pEU-E01-FLAG-ASC-PYD (#RDB19446)

A wheat germ cell-free expression plasmid of the N-terminal FLAG-tagged pyrin domain of ASC.

Clone info. A wheat germ cell-free expression plasmid of the N-terminal FLAG-tagged pyrin domain (PYD) of Apoptosis associated Speck like protein containing Caspase recruit domain (ASC). In cell-free reconstituted NLRP3 innflammasome system, the purified product can be an acceptor of amplified luminescence proximity homogeneous assay (ALPHA) process in vitro. See the FLAG-ASC-PYD protein construct at Fig.1 in Kaneko, N. et al. Sci. Rep. 10 (1): 13562, 2020.
Comment If you use a QIAGEN kit, it is recommended that you use the Plasmid Midi or Maxi Kit. The use of miniprep kits, including products from other companies, is not recommended. Expression was confirmed by the depositor with western blotting.
Vector backbone pEU-E01-FLAG-GW (plasmid)
Size of vector backbone 5.0 kb
Selectable markers Ampicillin
Growth conditions LB+Ampicillin, 30oC
Growth remarks Stbl2 is recommended E. coli for plamid amplication.
Gene/insert name human PYCARD cDNA
Depositor|Developer Masumoto, Junya |
 
Sequence (full) RDB19446hts01.seq checkAssembled from experimentally sequenced data.
Publication Kaneko, N., Sci. Rep. 10 (1): 13562 (2020). PMID 32782316. [link to RRC of NBRP]
Test sheet RDB19446C0p1-1.pdf 
 

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
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MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ]
Please visit Information of Request for Distribution.[link] 
Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested. (Kaneko, N. et al., Sci. Rep. 10 (1): 13562, 2020)
Remarks
Gateway® Expression clone. Please refer Life Technologies Corporation signs license agreement with RIKEN BRC.
提供案内 (日本国内) [open/close]

必要書類
提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
手続きの概要は、「提供申込みについて[link]」をご覧ください。
MTAに書く使用条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する(Kaneko, N. et al., Sci. Rep. 10 (1): 13562, 2020)。
備考
Gateway®テクノロジーを用いた発現クローンです。「Gateway®テクノロジーを用いたクローンの提供と寄託のご案内」をご覧ください。

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB19446 pEU-E01-FLAG-ASC-PYD DNA solution JPY 9,460 (not-for-profit academic purpose)
plus cost of shipping containers, dry ice (if required) and shipping charge plus licensing fee equivalent for Gateway® expression clone

How to cite this biological resource

Materials & Methods section:

The pEU-E01-FLAG-ASC-PYD was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB19446).

Reference section:

Kaneko, N., Kurata, M., Yamamoto, T., Shigemura, T., Agematsu, K., Yamazaki, T., Takeda, H., Sawasaki, T., Koga, T., Kawakami, A., Yachie, A., Migita, K., Yoshiura, K.I., Urano, T., Masumoto, J., KN3014, a piperidine-containing small compound, inhibits auto-secretion of IL-1β from PBMCs in a patient with Muckle-Wells syndrome. Sci. Rep. 10 (1): 13562 (2020). PMID 32782316. [link to RRC of NBRP]

Further references such as user reports and related articles (go to bottom)

References

Original, user report and related articles

original Kaneko, N., KN3014, a piperidine-containing small compound, inhibits auto-secretion of IL-1β from PBMCs in a patient with Muckle-Wells syndrome. Sci. Rep. 10 (1): 13562 (2020). PMID 32782316. [link to RRC of NBRP]
reference Kaneko, N., Poly (I:C) and hyaluronic acid directly interact with NLRP3, resulting in the assembly of NLRP3 and ASC in a cell-free system. Eur. J. Inflamm. 15 (2): 85-97 (2017). [link to Journal]