RDB14601 - pMRX-IP-GFP-LC3-RFP

Fluorescent probe to evaluate autophagic flux by the rat LC3 fused with GFP and RFP.

Distribution information

Catalog number	RDB14601
Resource name	pMRX-IP-GFP-LC3-RFP
Clone info.	Fluorescent probe to evaluate autophagic flux. Expression vector of rat microtubule-associated protein 1 light chain 3 beta (LC3) fused with GFP and RFP. This probe is cleaved by endogenous ATG4 proteases into equimolar amounts of GFP-LC3 and RFP. The GFP-LC3 is degraded by autophagy, while RFP remains in the cytosol as an internal control. Thus, autophagic flux can be estimated by calculating the GFP/RFP signal ratio. Please refer DISCUSSION section of Kaizuka, T., Mol. Cell 64, 835-849, 2016 for the difference between pMRX-IP-GFP-LC3-RFP-LC3deltaG and this clone. Expression was confirmed by the depositor with immunoblotting and fluorescent microscopy. Please refer Saitoh, T. et al., FEBS Lett., 532 (1-2): 45-51, (2002) for the construction of pMRX-IP vector.
Vector backbone	pMRX-IP (Plasmid)
Selectable markers	Amp^r (E. coli), Puro^r (mammalian cells).
Gene/insert name	Rat LC3 cDNA Aequorea victoria GFP cDNA Discosoma sp. RFP cDNA
Depositor\|Developer	Mizushima, Noboru \|

Other clones in our bank	Rat Map1lc3b (KEGG orthology K10435) \|

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.

Terms and conditions set forth by the DEPOSITOR	(1) BIOLOGICAL RESOURCE is limited to for academic research in non-profit organization. (2) The RECIPIENT of BIOLOGICAL RESOURCE shall obtain a prior written consent on use of it from the DEPOSITOR. (3) Any use of the BIOLOGICAL RESOURCE for profit purposes by a non-profit organization or any use of the BIOLOGICAL RESOURCE by a profit organization requires a separate license from the DEPOSITOR prior to distribution. (4) In publishing research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the original publication designated by the DEPOSITOR (Kaizuka, T. et al. Mol. Cell, 64 (4): 835-849, 2016) and an acknowledgment to the DEPOSITOR are requested. (5) The RECIPIENT must contact the DEPOSITOR in case of application for any patents or commercial use with the results from the use of BIOLOGICAL RESOURCE.
Additional terms and conditions for distribution	Regarding pMRX vector: In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literatures for pMX and pMRX vectors (Kitamura, T., Exp. Hematol. 31 (11): 1007-1014, 2003; Saitoh T., J. Biol. Chem. 278 (38): 36005-36012, 2003) is requested.
Ordering	Please visit Information of Request for Distribution.[link] UCSD's MTA is also required. Please visit Additional Forms section. Order form [Credit Card Payment] [Bank Transfer or Check Payment] Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word] Schedule A (GFP-A) [Link] Approval form by depositor [Word]
Remarks	((Additional form)) Approval Form(FormD) Remember that you will be working with samples containing infectious virus. UCSD's MTA is also required. Please visit Additional Forms section. Please use Schedule A (GFP-A) additionally. Please obtain written approval of the depositor.

Distribution information (domestic users)

[open/close]

提供条件	1. 非営利団体による学術研究に限る。2. 利用者は事前に寄託者の提供承諾書を得る。3. 当該リソースの非営利団体の営利目的での利用および営利団体の利用については、事前に寄託者との間で、別途使用許諾に係る契約の締結が必要である。4. 研究成果の公表にあたって寄託者の指定する文献(Kaizuka, T. et al. Mol. Cell, 64 (4): 835-849, 2016)を引用し、謝辞の表明を必要とする。5. 利用者が本件リソースを使用して得られた研究成果に基づき特許等の申請、及び事業活動を行う場合は、寄託者と別途協議を行う。
付加的提供条件	pMRXベクターについて：利用者は、研究成果の公表にあたって pMXならびにpMRXベクターの文献（Kitamura, T., Exp. Hematol. 31 (11): 1007-1014, 2003; Saitoh T., J. Biol. Chem. 278 (38): 36005-36012, 2003) の引用を必要とする。
提供依頼	手続きの詳細は、「提供申込みについて[link]」をご覧ください。別途、UCSDとのMTAも必要です。「申込みに必要な書式」をご覧ください。提供依頼書 [Word] 提供同意書 (MTA、非営利機関による非営利学術研究用)[Word] Schedule A（GFP-A, 英語版のみ） [Link] 寄託者による提供承諾書 [Word]
備考	【追加提供依頼書】承諾書(FormD) このリソースはウイルス粒子産生用のため、取扱いに注意が必要です。別途、UCSDとのMTAも必要です。「申込みに必要な書式」をご覧ください。 Schedule A（GFP-A, 英語版のみ）が追加で必要です。寄託者の書面による提供承諾が必要です。

References and tips

Sequence information

Catalog #	Resource name	Shipping form	Fee (non-profit org.)
RDB14601	pMRX-IP-GFP-LC3-RFP	DNA solution

original	Morishita, H., A new probe to measure autophagic flux in vitro and in vivo. Autophagy 13 (4): 757-758 (2017). PMID 28121224.
original	Kaizuka, T., An Autophagic Flux Probe that Releases an Internal Control. Mol Cell. 2016 Nov 17;64(4):835-849 (2016). PMID 27818143.

Featured content	Autophagy (English text)

RIKEN BRC has sequenced portions of this material for quality test.
Please review the QC test results indicated by check

icon as well as clone information before placing your order.

Primer: EGFP-C Sequence file: RDB14601_A7Gva.seq
//
Primer: CSII-CMV3' Sequence file: RDB14601_A7Gvb.seq
//
Primer: MMLV-pol_F2 Sequence file: RDB14601_A7Gvc.seq
//
Primer: EGFP-N Sequence file: RDB14601_A7Gvd.seq
//
Primer: pJB8-Forward Sequence file: RDB14601_A7Gve.seq
//
Primer: p15Aori_F Sequence file: RDB14601_A7Gvf.seq
//
Primer: pMXs-3LTRup_R2 Sequence file: RDB14601_A7Gvg.seq
//