Wild-type monomeric red fluorescent protein (mRFP) Rab5 (pmRFP-C3) construct (Vonderheit and Helenius, 2005 (link)) was a gift from Ari Helenius (ETH Zurich; Addgene plasmid 14437). The constitutively active mCherry-Rab5CA(Q79L) (PmCherry-C1; Addgene plasmid 35138) and dominant-negative mCherry-Rab5DN(S34N) (PmCherry-C1; Addgene plasmid 35139) were gifts from Sergio Grinstein (University of Toronto; Bohdanowicz et al., 2012 (link)). The DsRed-Rab7 WT (pDsRed-C1; Addgene plasmid 12661), DsRed-Rab9 WT (pDsRed-C1; Addgene plasmid 12677), and DsRed-Rab11 WT (pDsRed-C1; Addgene plasmid 12679) were gifts from Richard Pagano (Mayo Clinic; Choudhury et al., 2002 (link)).
Mcherry rab5ca q79l
Manufactured by Addgene
MCherry-Rab5CA(Q79L) is a fluorescent protein construct that consists of the constitutively active mutant form of the Rab5 GTPase, Rab5CA(Q79L), fused to the mCherry red fluorescent protein. Rab5 is a small GTPase that regulates early endosome formation and trafficking. The Q79L mutation renders Rab5 constitutively active, leading to the formation of enlarged endosomes.
Automatically generated - may contain errors
Lab products found in correlation
Mcherry rab5dn s34n, by Addgene (2 mentions)
Dsred rab9 wt, by Addgene (1 mentions)
Egfp rab7, by Addgene (1 mentions)
Nsg1 emerald, by Addgene (1 mentions)
Pbmn yfp dhfr dd, by Addgene (1 mentions)
Pcdna4 vps34 flag, by Addgene (1 mentions)
Ptag bfp c h rab5a c myc, by Addgene (1 mentions)
Kod plus mutagenesis kit, by Toyobo (1 mentions)
4 protocols using mcherry rab5ca q79l
1
Fluorescent Rab Protein Constructs
2
Fluorescent Protein-Tagged Rab GTPases
Check if the same lab product or an alternative is used in the 5 most similar protocols
Nsg1-Emerald:Addgene cat#54202, Michael Davidson lab; the mutation at residue 114 from D to G was corrected.
Nsg1-mCherry:mouse full length Nsg1 was cloned at the N-terminal of mCherry in pcDNA3 vector.
Nsg2-GFP:mouse full length Nsg2 was cloned into pcDNA-GFP by Genscript
mRFP-Rab5:Addgene cat#14437, Ari Helenius lab.
mCherry Rab5CA(Q79L) cat#35138, mCherry Rab5DN(S34N) cat#35139:Addgene, Sergio Grinstein lab.
EGFP-Rab7 cat#12605, EGFP-Rab7DN(T22N) cat#12660:Addgene, Richard Pagano labmTagBFP2-Rab5, cat#56417, Addgene, Michael Davidson lab;
mTagBFP2-Rab7:Michael Davidson lab.
GFP:Clontech
mCherry:Clontech.
+ Expand
Nsg1-Emerald:Addgene cat#54202, Michael Davidson lab; the mutation at residue 114 from D to G was corrected.
Nsg1-mCherry:mouse full length Nsg1 was cloned at the N-terminal of mCherry in pcDNA3 vector.
Nsg2-GFP:mouse full length Nsg2 was cloned into pcDNA-GFP by Genscript
mRFP-Rab5:Addgene cat#14437, Ari Helenius lab.
mCherry Rab5CA(Q79L) cat#35138, mCherry Rab5DN(S34N) cat#35139:Addgene, Sergio Grinstein lab.
EGFP-Rab7 cat#12605, EGFP-Rab7DN(T22N) cat#12660:Addgene, Richard Pagano labmTagBFP2-Rab5, cat#56417, Addgene, Michael Davidson lab;
mTagBFP2-Rab7:Michael Davidson lab.
GFP:Clontech
mCherry:Clontech.
3
Cloning and Tagging of Golgi Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cloning of B4GT-EGFP-HT2 was previously described (Serebrenik et al., 2018 (link)). B4GT-SNAP-HT2 was cloned by replacing the EGFP domain of B4GT-EGFP-HT2 using Gibson assembly (Gibson et al., 2009 (link)). ST6GAL1-EGFP-HT2 and MAN2A1-EGFP-HT2 were cloned by replacing the transmembrane and cytoplasmic domain of B4GT-EGFP-HT2 with the first 113 and 88 amino acids of ST6GAL1 (Homo sapiens) and MAN2A1 (Mus musculus), respectively, via USER cloning (Nour-Eldin et al., 2010 ). B4GT-EGFP-DHFR* was created by replacing HT2 from B4GT-EGFP-HT2 with the DHFR domain from pBMN YFP-DHFR(DD) (a gift from Thomas Wandless [Stanford University], Addgene plasmid #29326). mCherry-Rab5CA(Q79L) was a gift from Sergio Grinstein (Addgene plasmid #35138). RFP-Rab5a and B4GT-GFP were gifts from James Rothman (Yale University), from which B4GT-RFP was derived by replacing the GFP domain with RFP via USER cloning. B4GT-3xFM*-GFP was a gift from Adam Linstedt (Carnegie Mellon University).
4
Rab5-Mediated Vesicle Dynamics Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
pET-GST-R5BD-hRABEP1 was purchased from Vector Builder. Rab5a-pmCherryC1 was a gift from Christien Merrifield (Addgene plasmid #27679; http://n2t.net/addgene:27679 , accessed on 31 May 2019; RRID:Addgene_27679) [41 (link)]. pcDNA4-Vps34-Flag was from Qing Zhong (Addgene plasmid #24398; http://n2t.net/addgene:24398 , accessed on 12 November 2018; RRID:Addgene_24398) [42 (link)]. mCherry-Rab5CA(Q79L) was from Sergio Grinstein (Addgene plasmid #35138; http://n2t.net/addgene:35138 , accessed on 26 April 2018; RRID:Addgene_35138) [43 (link)]. TagRFP-T-EEA1 was from Silvia Corvera (Addgene plasmid #42635; http://n2t.net/addgene:42635 , accessed on 9 November 2017; RRID:Addgene_42635) [44 (link)]. pTag-BFP-C-h-Rab5a-c-Myc was from James Johnson (Addgene plasmid #79801; http://n2t.net/addgene:79801 , accessed on 9 November 2017; RRID:Addgene_79801) [45 (link)]. RFP-Rab5 140 and 180 mutants were generated with primers (5′-GCAGCAGCTGTTGACTTCCAGG-3′ and 5′-ATTTGCTAAGTCAGCTTTGTTTCCTGAC-3′) and (5′-GCAGCGCTGCCAAAGAATGAAC-3′ and 5′-AGCTATTGCCATAAATATTTCATTTACATTCATTG-3′), respectively, using a KOD-plus mutagenesis kit (Toyobo, Osaka, Japan).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!