B6.Cg-Fcgrttm1Dcr Tg(FCGRT)276Dcr mice deficient in mouse FcRn α-chain gene, but hemizygous transgenic for a human FcRn α-chain gene (mFcRn−/− huFcRn tg +/−, line 276), were used for the pharmacokinetic studies (38 (link)). A single dose of antibody was injected i.v. via the lateral tail vein at a dose level of 10 mg/kg. The mice were divided into three groups of six mice each to cover nine serum collection time points in total (at 0.08, 2, 8, 24, 48, 168, 336, 504, and 672 h post-dose). Each mouse was subjected twice to retroorbital bleeding, performed under light anesthesia with isoflurane (CP-Pharma GmbH); a third blood sample was collected at the time of euthanasia. Blood was collected into serum tubes (Microvette 500Z-Gel; Sarstedt). After 2 h incubation, samples were centrifuged for 3 min at 9,300 × g to obtain serum. After centrifugation, serum samples were stored frozen at −20 °C until analysis.
Microvette 500 z gel
Manufactured by Sarstedt
Sourced in Germany
The Microvette 500 Z-Gel is a laboratory equipment product manufactured by Sarstedt. It is a collection and storage tube designed for the collection and handling of small sample volumes. The product features a gel separation medium to facilitate the separation of serum or plasma from the cellular components of the sample.
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Lab products found in correlation
Isoflurane, by CP-Pharma (1 mentions)
Prism 7, by GraphPad (1 mentions)
Dexmedetomidine hydrochloride, by Orion Pharma (1 mentions)
Quantichrom iron assay kit, by BioAssay Systems (1 mentions)
Milliplex map kit, by Merck Group (1 mentions)
Ratlaps ctx 1 elisa kit, by Immunodiagnostic Systems (1 mentions)
9 protocols using microvette 500 z gel
1
Pharmacokinetics of Antibody in FcRn Mice
2
Quantifying Serum Gas6 Levels
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Peripheral blood serum was isolated using Microvette 500 Z-Gel (#20.1344, SARSTEDT). Samples were then analyzed using the Mouse Gas6 DuoSet ELISA (DY986: R&D Systems), according to the manufacturer's instructions. Results were plotted using GraphPad Prism 7 software.
3
Serum Collection from Immunized Mice
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Whole blood serum was collected from the immunized mice (n = 20), and non‐immunized TgN3R182C150 at 3 months (n = 9) and 7 months (n = 6). Briefly, mice were anesthetized intraperitoneally with avertin (240 mg/kg) and blood was then collected from the right ventricle and dispensed to a tube with serum gel with clotting activator (Microvette 500 Z‐Gel, Sarstedt). After centrifugation for 5 min at 10,000 g, the whole blood serum was collected, aliquoted, and stored at −80°C.
4
Testosterone Analysis in Mouse Serum
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Peripheral blood samples from all mice were collected at termination in 500 µL tubes containing serum gel with a clotting activator (Microvette 500 Z-Gel, Sarstedt, Numbrecht, Germany). The serum was extracted and stored at −80°C until use. Steroids were extracted from serum (200 µL) and concentrations of testosterone were analyzed by liquid chromatography-mass spectrometry as previously described (HPLC-MS; Acquity UPLC system and TQ-XS triple quadrupole mass spectrometer) (Ohlsson et al. 2022 (link)).
5
Comprehensive Mouse Tissue Sampling
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Mice were anesthetized with a mixture of ketamine (Richter Pharma)/dexmedetomidine hydrochloride (Orion Pharma) and euthanized by exsanguination followed by cervical dislocation. Peripheral blood was collected in 500 µL tubes containing serum gel with a clotting activator (Microvette 500 Z‐Gel; Sarstedt). Serum was extracted and stored at −80°C until use. Lung lavage was performed twice with 250 µL of ice‐cold PBS to collect bronchoalveolar lavage fluid (BALF). The liver was perfused by injection of 10 mL of PBS (room temperature) into the portal vein and the gall bladder was removed. Lungs, mediastinal lymph nodes (MLN), and livers were dissected, and weights were noted.
6
Serum Preparation and Analysis
7
Serum and Ascites Collection for Research
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To obtain the human serum, blood was withdrawn from healthy volunteers and collected into VenosafeTM tubes (6 mL) containing gel and clotting activator (Terumo EuropeTM, Leuven, Belgium) at Ghent University Hospital. Thereafter, the tubes were centrifuged (4000 g, 10 min, 20 °C) and serum (supernatant) was transferred into microvette ® 500 Z-Gel (SARSTEDT, Numbrecht, Germany) and subjected to centrifugation (10000 g, 5 min, 20 °C). The obtained serum was then stored at -20 °C. Furthermore, human ascites was obtained from patients with peritoneal carcinomatosis at the department of medical oncology, Ghent University Hospital, according to an approved procedure by the ethics committee of the Ghent University Hospital (EC no. 2013/589).
8
Serum and Bone Marrow Biomarker Quantification
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Peripheral blood samples were collected in clotting activator containing tubes (Microvette 500 Z‐Gel, Sarstedt, Nümbrecht, Germany) and serum was extracted. For BM plasma, femurs and tibias were dissected, ends were cut off, and bones were centrifuged in Eppendorf tubes for 30 seconds at 600g, 4°. The BM pellets were resuspended in 150 μL PBS per bone and collected in one tube for each mouse. After centrifugation for 2 minutes, at 600g, 4°, the supernatant was collected as BM plasma. The bone resorption marker C‐terminal type I collagen fragments was measured in serum using an ELISA RatLaps kit (CTX‐I, Immunodiagnostic Systems, East Boldon, UK) and procollagen type I N propeptide (PINP, Immunodiagnostic Systems) was analyzed as a serum marker of bone formation. The stromal cell factor C‐X‐C motif chemokine 12 (CXCL12) (R&D Systems, Minneapolis, MN, USA) was measured in serum and bone marrow plasma. All ELISA assays were performed according to the manufacturer's instructions.
9
In Vivo Experiments with C57BL/6 Mice
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In vivo experiments with C57BL/6 mice were performed at Experimental Pharmacology & Oncology Berlin-Buch (EPO, Germany) in accordance with the United Kingdom Co-ordinating Committee on Cancer Research (UKCCCR) regulations for the Welfare of Animals36 and of the German Animal Protection Law and approved by the local responsible authorities.
Compounds were injected s.c. into the scapular region of mice. The administration volume was adjusted corresponding to the body weight (10 mL/kg). Blood was collected by orbital sinus bleeding, and serum was extracted by centrifugation using Microvette 500 Z-Gel (Sarstedt) and transferred into Eppendorf tubes for storage at −20°C.
Compounds were injected s.c. into the scapular region of mice. The administration volume was adjusted corresponding to the body weight (10 mL/kg). Blood was collected by orbital sinus bleeding, and serum was extracted by centrifugation using Microvette 500 Z-Gel (Sarstedt) and transferred into Eppendorf tubes for storage at −20°C.
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