Animal studies were performed according to the guideline for animal protection and approved by the institutional animal care and use committee. Thalidomide was added to a fat and cholesterol enriched western type diet (ssniff TD 88137, ssniff Spezialdiäten GmbH, Soest, Germany) and administered with a mean daily dose of 200 mg/kg/day to male AL mice (n = 5, Charles Rivers Wiga, Sulzbach, Germany) beginning at the age of 6 weeks. Coeval male AL mice (n = 5) with the same western diet (WD) but without thalidomide served as controls. Chocolate taste was added to both diets to conceal potentially disgusting taste. Animals were housed in a specific pathogen free environment with access to food and water ad libitum and a 12/12 h day/night cycle. After 29 weeks of diet, i.e. at age of 35 weeks, the animals were euthanized with a fatal dose of inhaled isoflourane. The left ventricle was cannulated and injected with heparinized saline (10 ml of 0.9 % sodium chloride with 1,000 IU heparin) until the venous effluent from the right atrium was free from blood. A plumbiferous radiopaque polymer (Microfil® MV-122, Flow Tech, Carver, MA, USA) was injected into the left ventricle at a nominal pressure of 100 mmHg until it emerged from the inferior vena cava. After polymerization of the compound, the heart and entire aorta were removed and immersed in 4 % neutral buffered formalin.
Td88137
Manufactured by Ssniff
Sourced in Germany
The TD88137 is a laboratory instrument designed for precise measurement and analysis. Its core function is to perform high-accuracy quantitative assessments of various samples. The device utilizes advanced technology to ensure reliable and consistent results.
Automatically generated - may contain errors
Lab products found in correlation
Apoe mice, by Jackson ImmunoResearch (4 mentions)
Xylazine, by Bayer (3 mentions)
Ketamine, by Pfizer (3 mentions)
B6.129p2 lyz2tm1 cre ifo j, by Jackson ImmunoResearch (2 mentions)
C57bl 6j, by Janvier Labs (2 mentions)
Microfil mv 122, by Flow Tech (1 mentions)
Rosa26flox mt stop flox mg, by Jackson ImmunoResearch (1 mentions)
Tamoxifen, by Merck Group (1 mentions)
C57bl 6n, by Charles River Laboratories (1 mentions)
Phosphate buffered saline (pbs), by Lonza (1 mentions)
C57bl 6 wt, by Charles River Laboratories (1 mentions)
Accustain elastic stain, by Merck Group (1 mentions)
Arg1flox, by Jackson ImmunoResearch (1 mentions)
Apoe b6.129p2 apoetm1unc j, by Jackson ImmunoResearch (1 mentions)
22 protocols using td88137
1
Thalidomide Effects on Vascular Remodeling
2
Atherosclerosis Model in Transgenic Mice
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The Animal Ethics Committees of Karlsruhe and Darmstadt approved all mouse procedures (Protocol No. B2/1009). Generation of transgenic mouse lines on the C57BL6/N background was described previously: for smooth muscle myosin heavy chain transgenic mice (SMMHC-CreER T2 ) [26] and for monocytes/macrophages (LysM-Cre) [5] . These mice were bred on the ApoEdeficient mouse line [17] and then crossed with the double fluorescent reporter line Rosa26 flox-mT-stop-flox-mG (Jackson Lab, Stock 007576) reported by Ref. [16] . Mice were housed under a 12 h lightdark cycle with free access to food and water and under pathogenfree conditions. Cre recombinase was activated in male mice at 6e8 weeks of age with intraperitoneal injections of tamoxifen (1 mg/ mouse, Sigma T5648), one per day for 5 consecutive days. Five days after the last tamoxifen injection, mice were fed a high fat diet for 16 weeks to accelerate development of atherosclerotic lesions. Diet contained 21% butter fat and 1.5% cholesterol (Ssniff ® TD88137). Animals were euthanized by CO 2 under intraperitoneal anesthesia of ketamine (120 mg/kg, Pfizer, Germany) and xylazine (16 mg/kg, Bayer; Germany), and then perfused via the left ventricle with phosphate-buffered saline (PBS).
3
Intestinal Segment Analysis in Mice
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Eleven-week old male C57BL/6J mice were fed with Western-type diet (TD88137 mod., 21% fat, 0.2% cholesterol; Ssniff Spezialdiaeten GmbH, Soest, Germany) for 4 weeks. Thereafter, animals were fasted overnight (12 h) and gavaged with 120 µl of corn oil. Two hours post-gavage, mice were sacrificed by cervical dislocation, small intestines were dissected, briefly washed in PBS and cut in ∼ 3 cm long pieces. Luminal sides of each piece were scraped, scrapings were thoroughly mixed, split in 2 parts and used as stated below. The first 3 fractions were designated as duodenum, fractions 4-7 as jejunum, and the 4 most distal fractions as ileum.
4
High-fat Diet Impacts on C57Bl/6J Mice
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C57Bl/6J were used and had free access to food and water and were housed in climate and light controlled quiet rooms with a 12 h light–dark cycle. General well-being was assessed by daily inspections and monitoring of body weights (1–2/week). The experiments were approved by the local Ethics Committee for animal research (Darmstadt, Germany), adhered to the guidelines of the Society of Laboratory Animals (GV-SOLAS) and were in line with the ARRIVE guidelines and the European and German regulations for animal research. Male 24-week-old C57Bl/6J mice (n = 6) were fed with a high-fat Western-type diet (ssniff, Soest, Germany, diet TD88137) for 16 weeks. Control 22–23 weeks old C57Bl/6J mice (n = 6) were fed with standard food pellets in parallel. Mice were sacrificed by CO2 intoxication and cardiac puncture, and the liver and visceral fat were excised and immediately shock-frozen in liquid nitrogen. Tissue samples were stored at -80 °C until RNA extraction.
5
Dietary Intervention in Mice
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Age-matched male LAL-KO mice and their corresponding WT littermates [12 (link)] on a C57BL/6J background [16 (link)] were used for all experiments unless otherwise indicated. Mice had ad libitum access to water and food and were maintained under a 12 h light/12 h dark cycle in a temperature-controlled environment. Mice were fed a standard chow diet (Altromin 1324, Lage, Germany), after which the animals were challenged with a Western-type diet (WTD) (TD88137; 21% fat, 0.2% cholesterol; Ssniff Spezialdiaeten GmbH, Soest, Germany) for 2–6 weeks. All experiments were performed in accordance with the European Directive 2010/63/EU and approved by the Austrian Federal Ministry of Education, Science and Research (Vienna, Austria; BMWFW-66.010/0065-WF/V/3b/2015, BMWFW-66.010/0081-WF/V/3b/2017, BMBWF-66.010/0106-V/3b/2019; 2020-0.129.904).
6
Investigating Atherosclerosis in Siglec-G Deficient Mice
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All experimental protocols were approved by the institutional animal experimentation committee and the Austrian Ministry of Science. All mice were on a C57BL/6 background. Ldlr−/− and μMT mice were purchased originally from The Jackson Laboratories (Bar Harbor, Maine, USA). The generation of C57BL/6 Siglec-G−/− mice has been described elsewhere (Müller et al., 2015 (link)). Siglec-G−/− mice were further crossed with Ldlr−/− mice to obtain Ldlr−/−Siglec-G−/− and Ldlr−/−Siglec-G+/+ mice. For intervention studies, three cohorts of 12-week-old male Ldlr−/−Siglec-G−/− mice (n = 14) and Ldlr−/−Siglec-G+/+ (n = 16) littermate controls were fed an atherogenic diet containing 21% milk fat and 0.2% cholesterol (TD88137, Ssniff Spezialdiäten) for 8 weeks. Bone marrow transplantation studies were performed as previously described (Binder et al., 2004 (link), Sage et al., 2012 (link), Fillatreau et al., 2002 (link)) and as described in Supplemental Information .
7
Endothelium-Specific Signaling Pathways in Atherosclerosis
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All mice were backcrossed onto a C57BL/6N background (Charles River), and experiments were performed with littermates as controls. Male animals (8–12 weeks old) were used unless stated otherwise. Mice were housed under a 12-hour light/12-hour dark cycle, with free access to food and water and under specific pathogen–free conditions unless stated otherwise. The generation of inducible endothelium-specific Gαs–deficient mice (Tie2-CreERT2;Gnasfl/fl), endothelium-specific CALCRL–, and adrenomedullin-deficient mice (Tie2-CreERT2;Calcrlfl/fl and Tie2-CreERT2;Admfl/fl, respectively) was described previously (25 (link)). Cre-mediated recombination was induced by i.p. injection of tamoxifen (MilliporeSigma, T5648) dissolved in Miglyol 812 (1 mg per mouse per day) on 5 consecutive days.
For atherosclerosis studies, endothelium-specific Gαs– or CALCRL- deficient mice were crossed with the low-density lipoprotein receptor KO (Ldlr-KO) and were fed a high-fat diet for 14 weeks to induce development of atherosclerotic lesions. Diet contained 21% butterfat and 1.5% cholesterol (Ssniff, TD88137). For atherosclerosis studies in endothelium-specific adrenomedullin–deficient mice, mice were injected with 1 × 1011 vector genome copies (VG) of AAV-PCSK9 or AAV-Luc via the tail vein 3 days after the last tamoxifen injection.
For atherosclerosis studies, endothelium-specific Gαs– or CALCRL- deficient mice were crossed with the low-density lipoprotein receptor KO (Ldlr-KO) and were fed a high-fat diet for 14 weeks to induce development of atherosclerotic lesions. Diet contained 21% butterfat and 1.5% cholesterol (Ssniff, TD88137). For atherosclerosis studies in endothelium-specific adrenomedullin–deficient mice, mice were injected with 1 × 1011 vector genome copies (VG) of AAV-PCSK9 or AAV-Luc via the tail vein 3 days after the last tamoxifen injection.
8
Carotid Artery Ligation Model of Atherosclerosis
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Partial carotid artery ligation was performed as described before (34 (link)). In brief, 10 days after the last tamoxifen injection, mice were anesthetized by i.p. injection of ketamine (120 mg/kg, Pfizer) and xylazine (16 mg/kg, Bayer) and placed on a heated surgical pad. After hair removal, a midline cervical incision was made, and the left internal and external carotid arteries were exposed and partially ligated with 6.0 silk sutures (Serag-Wiessner), leaving the superior thyroid artery intact. Skin was sutured with absorbable 6.0 silk suture (CatGut), and animals were monitored until recovery in a chamber on a heating pad after the surgery. Animals were fed a high-fat diet for 2 weeks (Ssniff, TD88137), at which time their carotid arteries were harvested. To determine atherosclerotic lesions, left (partially ligated) and right (sham) carotid arteries were removed and fixed in 4% PFA overnight. Fixed vessels were embedded in paraffin. Serial sections (5 μm) were made through the entire carotid arteries, and a defined segment (1000–1500 μm from the ligature, unless otherwise stated) were stained with accustain elastic stain (MilliporeSigma) according to manufacturer’s instructions. Plaque area was calculated by subtracting the lumen area from the area circumscribed by the internal elastic lamina.
9
Metabolic Profiling of Ifrd1/2 Knockout Mice
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Age-matched (7-week-old) male Ifrd1-/- Ifrd2-/- and WT mice were caged individually and maintained up to 3 wk on a synthetic, HFD diet (TD.88137; Ssniff). Animals were weighted every fourth day between 08:00 and 10:00. Intestines, liver, muscles, and adipose tissue were collected for total RNA and protein isolation. Unfixed intestines were flushed with PBS using a syringe, embedded in Tissue-Tek (Sakura, 4583) and frozen in liquid nitrogen for immunohistochemical analysis.
10
Metabolic Effects of Cholesterol-Rich Diet in Mice
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This study was approved by the German Animal Studies Committee of Schleswig-Holstein and was conducted in compliance with international guidelines. Two groups of female mice, with genotypes Cyp17a1(d/d) × ApoE(d/d) and ApoE(d/d), were studied. The symbol “d” stands for deletion and is similar to KO or “−”. From the age of 10 weeks, mice were fed either a standard chow diet (chow) or a Western-type diet (WTD) containing 0.2% cholesterol and 21.2% fat (TD.88137; ssniff Spezialdiäten GmbH, Soest, Germany) for 8 weeks. The mice were maintained under controlled conditions of temperature (23 °C), humidity (40–60%), and lighting (12 h/12 h light/dark cycle). After 8 weeks of diet-feeding, the mice were euthanized using an overdose of isoflurane inhalation, followed by cervical dislocation, and then perfused with phosphate-buffered saline, pH 7.4 (Lonza, Cologne, Germany). Blood samples were collected to study the lipid profiles of the mice, as described previously [16 (link)]. The body mass and visceral fat mass were determined as previously described [16 (link)]. Finally, the caeca of the mice containing feces were collected, and tail biopsies were obtained for re-genotyping. All the samples collected were stored at −80 °C until further analysis.
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