C. elegans strains were cultured as previously described by Brenner [11 (link)]. Strain VS20 hjIs67[Patgl-1::atgl-1::GFP] [12 (link)] was obtained from CGC and subsequently crossed into CB1370 daf-2(e1370) and MR1000 daf-2(e1370); aak-1(tm1944); aak-2(ok524) strains. Strains MR1348 daf-2(e1370); rrEx226[sur-5p::GFP::ATGL-1::HA; rol-6(gf)] and MR1413 daf-2(e1370); rrEx239[syr-5p::GFP::ATGL-1(S303A)::HA; rol-6(gf)] are described in [9 (link)]. The strain atgl-1(tm3116) harbors a 423bp deletion of the atgl-1 gene and was obtained from National BioResource Project, Tokyo, Japan. Rabbit polyclonal antibody against ATGL-1 was raised using a synthetic peptide CTKRKVPDEPTTSKR (GenScript). Anti-PAR-5 antibody was a gift from Dr. Andy Golden. Anti-GFP (abcam ab290), anti-ubiquitin (Santa Cruz SC8017) and anti-P-14-3-3 (Cell Signaling #2981S) antibodies are available commercially.
Ctkrkvpdepttskr
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CTKRKVPDEPTTSKR is a peptide sequence. It is a synthetic peptide that can be used in various research applications.
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2 protocols using ctkrkvpdepttskr
1
C. elegans Strains and Antibodies
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C. elegans strains were cultured as previously described by Brenner [11 (link)]. Strain VS20 hjIs67[Patgl-1::atgl-1::GFP] [12 (link)] was obtained from CGC and subsequently crossed into CB1370 daf-2(e1370) and MR1000 daf-2(e1370); aak-1(tm1944); aak-2(ok524) strains. Strains MR1348 daf-2(e1370); rrEx226[sur-5p::GFP::ATGL-1::HA; rol-6(gf)] and MR1413 daf-2(e1370); rrEx239[syr-5p::GFP::ATGL-1(S303A)::HA; rol-6(gf)] are described in [9 (link)]. The strain atgl-1(tm3116) harbors a 423bp deletion of the atgl-1 gene and was obtained from National BioResource Project, Tokyo, Japan. Rabbit polyclonal antibody against ATGL-1 was raised using a synthetic peptide CTKRKVPDEPTTSKR (GenScript). Anti-PAR-5 antibody was a gift from Dr. Andy Golden. Anti-GFP (abcam ab290), anti-ubiquitin (Santa Cruz SC8017) and anti-P-14-3-3 (Cell Signaling #2981S) antibodies are available commercially.
+ Expand
C. elegans strains were cultured as previously described by Brenner [11 (link)]. Strain VS20 hjIs67[Patgl-1::atgl-1::GFP] [12 (link)] was obtained from CGC and subsequently crossed into CB1370 daf-2(e1370) and MR1000 daf-2(e1370); aak-1(tm1944); aak-2(ok524) strains. Strains MR1348 daf-2(e1370); rrEx226[sur-5p::GFP::ATGL-1::HA; rol-6(gf)] and MR1413 daf-2(e1370); rrEx239[syr-5p::GFP::ATGL-1(S303A)::HA; rol-6(gf)] are described in [9 (link)]. The strain atgl-1(tm3116) harbors a 423bp deletion of the atgl-1 gene and was obtained from National BioResource Project, Tokyo, Japan. Rabbit polyclonal antibody against ATGL-1 was raised using a synthetic peptide CTKRKVPDEPTTSKR (GenScript). Anti-PAR-5 antibody was a gift from Dr. Andy Golden. Anti-GFP (abcam ab290), anti-ubiquitin (Santa Cruz SC8017) and anti-P-14-3-3 (Cell Signaling #2981S) antibodies are available commercially.
2
C. elegans Strains and Antibody Generation
Check if the same lab product or an alternative is used in the 5 most similar protocols
C. elegans strains were cultured as previously described by Brenner [50 (link)]. The following alleles and strains were used: The strain RB2386 C37H5.3(ok3245) was obtained from CGC and subsequently crossed into CB1370 daf-2(e1370) and MR1000 daf-2(e1370); aak-1(tm1944); aak-2(ok524) strains. ok3245 bears a 800bp deletion that occupies almost two thirds of the gene from the C-terminal, therefore, the allele is predicted to be null. C37H5.3 DNA and its upstream ~800bp (considered as its own endogenous promoter) was amplified by PCR and subsequently cloned into pPD95.77 GFP vector to generate pMR613 (Pcgi-58::cgi-58::GFP). Extrachromosomal arrays of pMR613 was generated by standard microinjection into CB1370 daf-2(e1370) animals using rol-6 cDNA rescue fragment as co-injection marker. Strain VS20 hjIs67[Patgl-1::atgl-1::GFP] (12) was obtained from CGC and subsequently crossed into CB1370 daf-2(e1370) and MR1000 daf-2(e1370); aak-1(tm1944); aak-2(ok524) strains.
Rabbit polyclonal antibody against ATGL-1 was raised using a synthetic peptide CTKRKVPDEPTTSKR (GenScript). Rabbit polyclonal antibody against GFP was raised by McGill Animal Resources Center. WT and CGI-58-/- mouse embryonic fibroblasts (MEFs) were gifts from Dr. Rudolf Zechner and were cultured according to their recommendation. Mouse polyclonal antibody against Actin was purchased from Abcam (ab14128).
+ Expand
C. elegans strains were cultured as previously described by Brenner [50 (link)]. The following alleles and strains were used: The strain RB2386 C37H5.3(ok3245) was obtained from CGC and subsequently crossed into CB1370 daf-2(e1370) and MR1000 daf-2(e1370); aak-1(tm1944); aak-2(ok524) strains. ok3245 bears a 800bp deletion that occupies almost two thirds of the gene from the C-terminal, therefore, the allele is predicted to be null. C37H5.3 DNA and its upstream ~800bp (considered as its own endogenous promoter) was amplified by PCR and subsequently cloned into pPD95.77 GFP vector to generate pMR613 (Pcgi-58::cgi-58::GFP). Extrachromosomal arrays of pMR613 was generated by standard microinjection into CB1370 daf-2(e1370) animals using rol-6 cDNA rescue fragment as co-injection marker. Strain VS20 hjIs67[Patgl-1::atgl-1::GFP] (12) was obtained from CGC and subsequently crossed into CB1370 daf-2(e1370) and MR1000 daf-2(e1370); aak-1(tm1944); aak-2(ok524) strains.
Rabbit polyclonal antibody against ATGL-1 was raised using a synthetic peptide CTKRKVPDEPTTSKR (GenScript). Rabbit polyclonal antibody against GFP was raised by McGill Animal Resources Center. WT and CGI-58-/- mouse embryonic fibroblasts (MEFs) were gifts from Dr. Rudolf Zechner and were cultured according to their recommendation. Mouse polyclonal antibody against Actin was purchased from Abcam (ab14128).
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