W1118

Manufactured by Bloomington Drosophila Stock Center

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W1118 is a wild-type Drosophila melanogaster strain commonly used as a genetic background. It serves as a standard reference strain for various experimental studies in the field of Drosophila research.

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Wild type w1118 (3 citations) W1118 strain (3 citations) Wild-type w1118 flies (2 citations) Yw and w1118 strains (2 citations) Standard laboratory strains w1118 (2 citations) Isogenic w1118 (2 citations) D. melanogaster strain w1118 (2 citations) W1118 line (2 citations) Drosophila melanogaster w1118 (2 citations) W1118 [3605]; (2 citations)

165 protocols using w1118

Fly Strain Rearing and Genotyping

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The fly strains and genotypes used in this manuscript are listed (Supplementary Data 8). All flies were reared on standard cornmeal–yeast–sucrose medium and kept in light/dark (LD) cycles at 25 °C. Food were accessible ad libitum. For propionylomic study, male w¹¹¹⁸ (Bloomington Stock Center, BL3605) flies were crossed with y¹,w^* and y¹,w^*,per^{0,25 (link)}. For RNA-seq, ChIP-seq and metabolomics studies, male w¹¹¹⁸ and H2BK17A flies were used. For the rest of the study, H2BK17A was backcrossed onto the isogenic w¹¹¹⁸ background (Bloomington Stock Center, #5905) for five generations and male flies were used along with isogenic w¹¹¹⁸ as control.
Drosophila S2 cells used in this study is a kind gift from Dr. Xi Zhou (Wuhan University). Human AC16 cells is a kind gift from Dr. Chengqi Xu (HUST). NIH 3T3, U2OS and HEK293 cells used in this study were from ATCC.

Shui K., Wang C., Zhang X., Ma S., Li Q., Ning W., Zhang W., Chen M., Peng D., Hu H., Fang Z., Guo A., Gao G., Ye M., Zhang L, & Xue Y. (2023). Small-sample learning reveals propionylation in determining global protein homeostasis. Nature Communications, 14, 2813.

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Drosophila Strains for Lipid Metabolism

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The following fly strains from the Bloomington Drosophila Stock Center were used in this study: CS (#64349), w¹¹¹⁸ (#3605), Oregon-R, y¹_,v¹;UAS-bmm-RNAi (#25926), y¹_,v¹;attP40 (#36304), w¹¹¹⁸;UAS-nGFP (#4775); UAS-mCD8::GFP (#5130). The following fly strains from the Vienna Drosophila Resource Center were used in this study: UAS-bmm-RNAi (#37880), UAS-bmm-RNAi (#37877). We obtained the bmm¹ mutants and bmm^rev control strain as a kind gift from R. Kühnlein [32 (link)]; CMW flies, Mel c2.2, and Mel c2.3 (wild-caught isofemale lines) as a kind gift from I. Dworkin; and UAS-LD-GFP flies from M. Welte. We used the following stocks for tissue-specific overexpression of the UAS-bmm-RNAi transgene: da-GAL4 (ubiquitous), cg-GAL4 (abdominal fat body), r4-GAL4 (abdominal fat body) elav-GAL4 (postmitotic neurons), nSyb-GAL4 (neurons), repo-GAL4 (glia), Mex-GAL4 (intestinal enterocytes), c587-GAL4 (somatic cells of the gonad), tj-GAL4 (somatic cells of the gonad), and dMef2-GAL4 (muscle cells).

Wat L.W., Chao C., Bartlett R., Buchanan J.L., Millington J.W., Chih H.J., Chowdhury Z.S., Biswas P., Huang V., Shin L.J., Wang L.C., Gauthier M.P., Barone M.C., Montooth K.L., Welte M.A, & Rideout E.J. (2020). A role for triglyceride lipase brummer in the regulation of sex differences in Drosophila fat storage and breakdown. PLoS Biology, 18(1), e3000595.

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Drosophila Strain Maintenance Protocol

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The following Drosophila strains were used in this study: esg-GFP/CyO and UAS-lacZ lines were kindly provided by Dr. Allan Spradling; esg^ts-Gal4 line was generously gifted from Dr. Benjamin Ohlstein; w¹¹¹⁸ (BDSC #3605), Canton-S (BDSC #64349), UAS-bsk^DN (BDSC #6409) and UAS-EGFR^DN (BDSC #5364) were obtained from the Bloomington Drosophila Stock Center. All flies used in this study were mated females unless otherwise mentioned.
Flies were maintained on standard cornmeal-agar medium (the recipe is: 80 g sucrose, 50 g cornmeal, 20 g glucose, 18.75 g yeast, 5 g agar, 30 mL propionic acid, and 1 L water) at 25°C and 60% humidity under a 12/12 h light/dark cycle. Gene overexpression driven by the esg^ts-Gal4 Drosophila line was repressed at 18°C and activated at 29°C.

Yan L., Zhou J., Yuan L., Ye J., Zhao X., Ren G, & Chen H. (2023). Silibinin alleviates intestinal inflammation via inhibiting JNK signaling in Drosophila. Frontiers in Pharmacology, 14, 1246960.

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Drosophila Fly Lines for Research

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The following Drosophila lines were used in the current study: w¹¹¹⁸ (BSC#9505, Bloomington Drosophila Stock Center, Bloomington, IN), Ore-R (wild type), w; hdc^Δ84/TM3, Kr>GFP[30] , lz-Gal4, UAS-GFP; +; + (a gift from Bruno Lemaitre, Lausanne, Switzerland) [32] (link), l(3)mbn¹/TM6 Tb[28] (link), and a homozygous hop^Tum (BSC#8492) line generated by Dr. Gábor Csordás (BRC, Szeged, Hungary). The flies were grown on a standard cornmeal-yeast substrate at 25 °C.

Balog J.Á., Honti V., Kurucz É., Kari B., Puskás L.G., Andó I, & Szebeni G.J. (2021). Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry. Genomics, Proteomics & Bioinformatics, 19(2), 243-252.

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Drosophila Rearing and Handling Protocol

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All flies were reared on standard medium at 25 C and approximately 65% humidity under a 12 h light/12 h dark cycle. Flies lines used in this study were as follows: esg-Gal4, UAS-GFP was kindly provided by Dr. Lihua Jin, Northeast Forestry University; 10×STAT92E-GFP and gstD1-GFP lines were gifts from Dr Fengwei Yu, Temasek Life Sciences Laboratory, National University of Singapore. w¹¹¹⁸ (#5905) were obtained from the Bloomington Drosophila stock center (BDSC; Indiana, United States). 3–5 day old adult female or male were collected using light CO₂ anesthesia and allowed to recover for 2 days before further experimentation.

Yang S., Li X., Xiu M., Dai Y., Wan S., Shi Y., Liu Y, & He J. (2022). Flos puerariae ameliorates the intestinal inflammation of Drosophila via modulating the Nrf2/Keap1, JAK-STAT and Wnt signaling. Frontiers in Pharmacology, 13, 893758.

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Drosophila Genetic Manipulation Techniques

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All fly stocks were maintained at 25°C on standard fly food medium. The following fly stocks were used: w¹¹¹⁸ [3605, Bloomington Drosophila Stock Center (BDSC)], GD control [60000, Vienna Drosophila Resource Center (VDRC)] and TRIP control (36303, BDSC) as control flies; and DMef2-Gal4 (Ranganayakulu et al., 1996 (link)) and Hand^4.2-Gal4 (Han and Olson, 2005 (link)) as driver lines. For CG14964, Df(3L)BSC672 (26524, BDSC) and CRIMIC line CG14964^{CR01157-TG4.1} (#81199, BDSC) were used; and for CG14964 RNAi GD (43603, VDRC) and TRIP3 (65245, BDSC) were used. For Mhc, we used Mhc¹ mutant (O'Donnell and Bernstein, 1988 (link)) and Mhc^YD0783 (50881, BDSC).

Auxerre-Plantié E., Nielsen T., Grunert M., Olejniczak O., Perrot A., Özcelik C., Harries D., Matinmehr F., Dos Remedios C., Mühlfeld C., Kraft T., Bodmer R., Vogler G, & Sperling S.R. (2020). Identification of MYOM2 as a candidate gene in hypertrophic cardiomyopathy and Tetralogy of Fallot, and its functional evaluation in the Drosophila heart. Disease Models & Mechanisms, 13(12), dmm045377.

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Drosophila Genetics and Fly Food Preparation

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Flies were cultured on yeast/molasses-based standard fly food (recipe: 10L H2O, 138g agar, 220g molasses, 750g malt extract, 180 dry yeast, 800g corn flour, 100g soy flour, 62.5ml propionic acid, 20g Methyl 4-Hydroxybenzoate, and 72ml ethanol). For GeneSwitch experiments, 86 mg RU486 was additionally dissolved in the ethanol for 200 μM final concentration. Flies were maintained at 25°C with a 12h light/dark cycle and 65% humidity. All flies used in the experiments reported were females.
Fly lines HmlΔ::Gal4, LSP2::Gal4, Elav::Gal4 and W¹¹¹⁸ were provided by the Bloomington Drosophila Stock Center. RNAi line MANFRNAiv12834 (UAS::MANFRNAi) was obtained from the Vienna Drosophila RNAi Center. These construct has no predicted off-targets. MANFRNAiv12834 was combined with a transgene driving Dicer overexpression (UAS::DICER2) to achieve efficient MANF knock-down^{19 (link)}.
We received the following lines as gifts: UAS::MANF, (T. Heino); NP1::Gal4 (D. Ferrandon); PPL::Gal4 (M. Pankratz); Upd3::Gal4 (N. Perrimon); C7::Gal4 (M. Uhlivora); UAS::Upd2 (M. Zeidler); UAS::Upd3 (N. Buchon); 2xStat::GFP (E. Bach).

Sousa-Victor P., Neves J., Cedron-Craft W., Ventura P.B., Liao C.Y., Riley R.R., Soifer I., van Bruggen N., Kolumam G.A., Villeda S.A., Lamba D.A, & Jasper H. (2019). MANF regulates metabolic and immune homeostasis in ageing and protects against liver damage. Nature metabolism, 1(2), 276-290.

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Drosophila Genetic Manipulation Protocols

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Standard methods were used for propagating fly stocks. For all experiments, embryos and larvae were raised at 25 °C, unless otherwise noted. The following lines were used: CQ2-GAL4 (Bloomington stock center [BL] 7468), OK6-GAL4 (BL 64199), hsFLP; UAS(FRT.stop)myr::smGdP-HA, UAS(FRT.stop)myr::smGdP-V5-THS UAS(FRT.stop)myr::smGdP-FLAG (BL 64085), UAS-myr-GFP (BL 32198), UAS-nls-GFP (BL 6452), UAS-Hb; UAS-HB/TM2 (BL 8504), w1118 (BL 36005), MHC-CD8-GCaMP6f-Sh (BL 67739) ac:VP16, gsb:v8v (aka NB7-1-GAL4, gift of M. Kohwi), VGlut-lexA/cyo (BL 60314), lexA(stop.FRT)mCD8.GFP (BL 57588), UAS-VP16::Hb/-;UAS-VP16::Hb (gift of C. Doe), UAS-svp1 1.12 (gift of M. Kowhi), Engrailed-GAL4 (BL 1973) (gift of M. Kowhi), UAS-hid,rpr (Zhou et al., 1997 (link)).

Meng J.L., Marshall Z.D., Lobb-Rabe M, & Heckscher E.S. (2019). How prolonged expression of Hunchback, a temporal transcription factor, re-wires locomotor circuits. eLife, 8, e46089.

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Genetic Manipulation of Antioxidant Capacity in Drosophila

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Flies were separated by sex and maintained in different vials on a standard sugar-yeast medium at constant temperature (25°C) and humidity (60%) in a 12:12 h light-dark cycle in Binder KBF720-ICH (Binder, Germany) climate chamber. The following strains were used: UAS-Gclc (provided by Dr. William C. Orr, Southern Methodist University) and Appl-GAL4 (#32040, Bloomington Drosophila Stock Center). They were backcrossed with w¹¹¹⁸ (#3605, Bloomington Drosophila Stock Center, USA) 6–8 times to equilibrate the genetic background. To activate the Gclc constitutive overexpression in the nervous system, the UAS-Gclc was crossed with Appl-GAL4 driver. Newly enclosed progenies were maintained for one, four, and 6 weeks before dissection. The same aged flies from the parental UAS-Gclc line were used as a contro control.

Moskalev A., Guvatova Z., Shaposhnikov M., Lashmanova E., Proshkina E., Koval L., Zhavoronkov A., Krasnov G, & Kudryavtseva A. (2019). The Neuronal Overexpression of Gclc in Drosophila melanogaster Induces Life Extension With Longevity-Associated Transcriptomic Changes in the Thorax. Frontiers in Genetics, 10, 149.

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Drosophila Neuronal Culture Protocols

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Flies were cultured on standard soft media seeded with live yeast at 25 °C unless otherwise indicated. w¹¹¹⁸, elav-Gal4, 24B-Gal4 and unc-104^d11024 were obtained from the Bloomington Drosophila Stock Center. GluRIIA-mRFP24 (link) and GluRIIB-GFP26 (link) with endogenous promoters were obtained from Stephan Sigrist (FU Berlin). UAS-unc-104-mcherry was a generous gift from Thomas Schwarz (Harvard University).

Zhang Y.V., Hannan S.B., Kern J.V., Stanchev D.T., Koç B., Jahn T.R, & Rasse T.M. (2017). The KIF1A homolog Unc-104 is important for spontaneous release, postsynaptic density maturation and perisynaptic scaffold organization. Scientific Reports, 7, 38172.

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