Topgal

Manufactured by Jackson ImmunoResearch

Topgal is a proprietary fluorescent labeling reagent developed by Jackson ImmunoResearch. It is designed for conjugating antibodies and other proteins to create fluorescent conjugates for use in a variety of immunoassay and cellular imaging applications.

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5 protocols using topgal

Generating Genetically Modified Mouse Models

Cited 1 time

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Dhcr7^−/− mice^{85 (link)} were a gift from Dr. Forbes D. Porter (The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA). Insig1^F/F;Insig2^−/− (The Jackson Laboratory, #005939)^{12 (link)} and Wnt1-Cre2 (The Jackson Laboratory, #022501)^{22 (link)} mice were obtained from The Jackson Laboratory and crossed to generate Insig1/2 cKO mice. Gli1-LacZ mice (The Jackson Laboratory, #008211)^{39 (link)} were obtained from The Jackson Laboratory and crossed with Dhcr7^+/− and Wnt1-Cre2;Insig1^F/+;Insig2^−/− mice in order to generate Dhcr7^−/−;Gli1-LacZ, Dhcr7^+/+;Gli1-LacZ, Insig1/2 cKO;Gli1-LacZ, and Insig1^F/F;Insig2^−/−;Gli1-LacZ mice. Topgal (The Jackson Laboratory, #004623)^{86 (link)} and Axin2^LacZ/+ (The Jackson Laboratory, #009120)^{87 (link)} mice were obtained from The Jackson Laboratory and crossed with Dhcr7^−/− mouse line to generate Dhcr7^−/−;Topgal and Dhcr7^+/+;Topgal, Dhcr7^−/−;Axin2^LacZ/+, and Dhcr7^+/+;Axin2^LacZ/+ mice. Genotyping was performed using PCR primers, as previously described.^{12 (link),22 (link),85 (link)} Pregnant females were treated with simvastatin (S6196; Sigma-Aldrich) at a dose of 10 mg/kg⁻¹ body weight (BW) from E12.5 to E18.5, or from day 7 to day 42, administered by intraperitoneal injection.

Suzuki A., Ogata K., Yoshioka H., Shim J., Wassif C.A., Porter F.D, & Iwata J. (2020). Disruption of Dhcr7 and Insig1/2 in cholesterol metabolism causes defects in bone formation and homeostasis through primary cilium formation. Bone Research, 8, 1.

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Conditional Knockout Mice for Lineage Tracing

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All animal experiments were authorized by the Animal Care and Use Committee of The University of Tokyo. We also complied with all relevant ethical regulations. In each experiment, we compared the genotypes of littermates maintained in a C57BL/6J background. TOPGAL, Prg4-Cre^ERT2, Rosa26-tdTomato (Ai14), and Ctnnb1-flox mice were obtained from The Jackson Laboratory (Bar Harbor, ME) [22 (link)–24 (link)]. Ctnnb1-ex3-flox mice were generated as previously described [25 (link)].

Xuan F., Yano F., Mori D., Chijimatsu R., Maenohara Y., Nakamoto H., Mori Y., Makii Y., Oichi T., Taketo M.M., Hojo H., Ohba S., Chung U.I., Tanaka S, & Saito T. (2019). Wnt/β-catenin signaling contributes to articular cartilage homeostasis through lubricin induction in the superficial zone. Arthritis Research & Therapy, 21, 247.

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Transgenic mouse strain generation

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The generation and genotyping of the following mouse strains have been described: K14-Eda [26 (link)], IκBαΔN [19 (link)], Eda null (Tabby) (Jackson Laboratories; stock no. 000314), TOP-gal (Jackson laboratories; stock no. 004623), K17-GFP, Foxi3-deficient, and NF-κB reporter mice [24 (link), 47 (link), 71 (link), 72 (link)]. K14-Eda, Foxi3-deficient, K17-GFP, and NF-κB rep mice were maintained on the C57Bl/6 background. IκBαΔN mice were bred in the C57BL/6 or a mixed C57BL/6 and FVB background. Eda null and TOP-gal mice were on B6CBA and NMRI backgrounds, respectively. The appearance of a vaginal plug was considered the embryonic day (E) 0.5. The age of the embryos were further staged according to the limb morphogenesis [73 (link)] and other external criteria.

Voutilainen M., Lindfors P.H., Trela E., Lönnblad D., Shirokova V., Elo T., Rysti E., Schmidt-Ullrich R., Schneider P, & Mikkola M.L. (2015). Ectodysplasin/NF-κB Promotes Mammary Cell Fate via Wnt/β-catenin Pathway. PLoS Genetics, 11(11), e1005676.

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Conditional Gene Targeting in Mice

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The Lrp6^flox mice for conditional gene-targeting analyses have previously been described (Zhou et al., 2010 (link)). The Pax3^Cre knock-in mice (Engleka et al., 2005 (link)), Rosa26-lacZ mice (Soriano, 1999 (link)), and canonical Wnt/β-catenin signaling reporter lines of BATgal (Maretto et al., 2003 (link)) and TOPgal (DasGupta and Fuchs, 1999 (link)) mice donated by different investigators were obtained through The Jackson Laboratory. Ctnnb1^flox(ex3) mice (MGI:1858008, gift from M. Taketo, Kyoto University, Kyoto, Japan) (Harada et al., 1999 (link)) were used for genetic rescue by conditional activation of β-catenin. These mouse strains were maintained on a C57BL/6J or a mixed B6;129 background. All mice were housed in the vivarium at the University of California, Davis (Davis, CA, USA). Pregnant, timed-mated mice were euthanized prior to cesarean section. Noon of the conception day was designated as E0.5. All research procedures using mice were approved by the University of California, Davis Animal Care and Use Committee and conformed to National Institutes of Health guidelines.

Zhao T., McMahon M., Reynolds K., Saha S.K., Stokes A, & Zhou C.J. (2022). The role of Lrp6-mediated Wnt/β-catenin signaling in the development and intervention of spinal neural tube defects in mice. Disease Models & Mechanisms, 15(6), dmm049517.

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Conditional Targeted Gpr177 Mouse Line

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The construction of conditional targeted Gpr177 mouse line, the Gpr177-Fx (Gpr177^fx/fx), has been described previously (Fu et al., 2009 (link)). The conditional targeted Gpr177 line was maintained in B6/C57 background as homozygous. Foxg1-Cre mouse was used to induce the tissue specific deletion of Gpr177 in the facial ectoderm during embryonic facial development. Briefly, Gpr177^fx/fx mice were crossed with Foxg1-Cre to obtain Gpr177^{(fx/+)Foxg1-Cre} mice which were back-crossed with Gpr177^fx/fx for Gpr177 conditional knockout mice. Wild-type or Gpr177^{(fx/+)Foxg1-Cre} littermates were used as control embryos. Mouse lines of R26R reporter, TOPGAL, Foxg1-Cre, and Wnt1-Cre were purchased from the Jackson Laboratory, Maine. All animal experimental protocols were approved by the Animal Users Committee of Hangzhou Normal University, China.

Zhu X.J., Liu Y., Yuan X., Wang M., Zhao W., Yang X., Zhang X., Hsu W., Qiu M., Zhang Z, & Zhang Z. (2016). Ectodermal Wnt Controls Nasal Pit Morphogenesis Through Modulation of the BMP/FGF/JNK Signaling Axis. Developmental dynamics : an official publication of the American Association of Anatomists, 245(3), 414-426.

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