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Please review the QC test results indicated by check icon below as well as clone information before placing your order.

pMRX-IP-GFP-LC3-RFP

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

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) |

Distribution information

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. Please contact us. UCSD's MTA is also required.
Order form [Credit Card Payment]  [Bank Transfer or Check Payment]
Material Transfer Agreement (MTA) [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.
Please contact us. UCSD's MTA is also required.
Please use Schedule A (GFP-A) additionally.
Please obtain written approval of the depositor.
Distribution information (domestic users) [open/close]


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

Ordering Information [in Japanese] [in English]
Please review the QC test results indicated by check icon below as well as clone information before placing your order.

References and tips

Electronic file

Original reference

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.

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

Featured content

Featured content Autophagy (English text)

Sequence information

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.

Test sheet RDB14601_A7Bap1_2.pdf check

Nucleotide sequence of a portion of this resource (if available).

Primer: EGFP-C
Sequence file: RDB14601_A7Baa.seq check
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Primer: CSII-CMV3'
Sequence file: RDB14601_A7Bab.seq check
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Primer: MMLV-pol_F2
Sequence file: RDB14601_A7Bac.seq check
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Primer: EGFP-N
Sequence file: RDB14601_A7Bad.seq check
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Primer: pJB8-Forward
Sequence file: RDB14601_A7Bae.seq check
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Primer: p15Aori_F
Sequence file: RDB14601_A7Baf.seq check
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Primer: pMXs-3LTRup_R2
Sequence file: RDB14601_A7Bag.seq check
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Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

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.
reference Nguyen, T.B., DGAT1-Dependent Lipid Droplet Biogenesis Protects Mitochondrial Function during Starvation-Induced Autophagy. Dev. Cell 42 (1): 9-21 (2017). PMID 28697336.
reference Tamura, N., Differential requirement for ATG2A domains for localization to autophagic membranes and lipid droplets. FEBS Lett. 591 (23): 3819-3830 (2017). PMID 29113029.

2020.07.02

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