A1048301

Manufactured by Thermo Fisher Scientific

Sourced in United States

The A1048301 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed to perform a core function within a laboratory setting, but a detailed description while maintaining an unbiased and factual approach is not available.

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Lab products found in correlation

47 protocols using a1048301

Subcutaneous Tumor Xenograft Assay

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1 × 10⁶ fibroblasts of each condition were suspended in 100 µL of FGM-2 bulletkit medium (CC-3132, Lonza) containing 50% type I collagen (A10483-01, Gibco). The suspensions were injected subcutaneously in the flank of CB17-SCID mice. For positive control, 1 × 10⁶ HT1080 cells suspended in 100 µL of DMEM containing 50% type I collagen (A10483-01, Gibco) were also injected subcutaneously in the flank of CB17-SCID mice. 12 mice were injected with nonirradiated NHDFs, 22 mice with PSNE cells, and 10 mice with HT1080 cells.

Goy E., Tomezak M., Facchin C., Martin N., Bouchaert E., Benoit J., de Schutter C., Nassour J., Saas L., Drullion C., Brodin P.M., Vandeputte A., Molendi-Coste O., Pineau L., Goormachtigh G., Pluquet O., Pourtier A., Cleri F., Lartigau E., Penel N, & Abbadie C. (2022). The out-of-field dose in radiation therapy induces delayed tumorigenesis by senescence evasion. eLife, 11, e67190.

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HUVEC Culture and Collagen Coating

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HUVECs were obtained from Lonza (Lot #0000636514). 3 days prior to system assembly HUVECs were thawed and plated. Rat tail collagen (Thermo Scientific cat #A1048301) was used to coat coverslips at a concentration of 500 µg/ml, with a seeding density of 4,500 cells/coverslip and maintained in vendor specific medium.

Rupar M.J., Sasserath T., Smith E., Comiter B., Sriram N., Long C.J., McAleer C.W, & Hickman J.J. (2023). Development of a human malaria-on-a-chip disease model for drug efficacy and off-target toxicity evaluation. Scientific Reports, 13, 10509.

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Collagen-based VSMC Contraction Assay

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Collagen gel-based assays were used to perform VSMC contraction studies according to previously reported methods.^{15 (link)} Collagen lattice at a final concentration of 1 mg/mL was prepared by mixing type-I collagen (ThermoFisher, A1048301), Vasculife ® SMC Smooth Muscle Cell Medium, Dulbecco’s Phosphate Buffered Saline (Invitrogen). 450 μL of collagen solution was added to 50 μL of VSMC suspension (4×10⁴ cells per well). Each 500 μL collagen/cell mixture was added to one well in a 96 well plate and allowed to polymerize for 30 minutes at 37 °C in a CO₂ incubator. After 30 minutes at 37 °C, 140 μL of VSMC media (2% FBS) was added to each well. Cells were incubated for 48 hours before imaging. Thirty minutes before imaging, collagen gels were released from the sides of the well with a 10 μL pipet tip and the media was replaced with VSMC media (10% FBS). The gel were imaged with a ThermoFisher EVOS FL Auto. Gel area was determined using ImageJ.

Koenig S.N., Cavus O., Williams J., Bernier M., Tonniges J., Sucharski H., Dew T., Akel M., Baker P., Madiai F., De Giorgi F., Scietti L., Faravelli S., Forneris F., Mohler P.J, & Bradley E.A. (2021). New Mechanistic Insights to PLOD1-mediated Human Vascular Disease. Translational research : the journal of laboratory and clinical medicine, 239, 1-17.

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Glass-PDMS Neuromuscular Junction Chambers

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NMJ chambers were constructed from glass bonded to polydimethylsiloxane (PDMS) casts, as described in our previous publications [12 (link),46 (link)]. Briefly, PDMS mold cut-outs were allowed to leach in 70% isopropyl alcohol overnight, after which they were rinsed in sterile water. Chambers were made by firmly pressing down sterilized and dry PDMS cut-outs on plasma-cleaned glass coverslips. The skeletal muscle and motoneuron sides of the chamber were coated with collagen I (ThermoFisher A1048301, Waltham, MA, USA) (100 µL Col I, 4.9 mls 1x PBS, 6 µL acetic acid) and Laminin (ThermoFisher 23017015, Waltham, MA, USA) (3 µg/mL in 1x phosphate buffered saline (PBS)), respectively, and allowed to incubate overnight at 4 °C prior to cell plating. Assembled chambers were brought to room temperature the next day before rinsing either side of each chamber with 1x PBS in preparation for cell plating.

Badu-Mensah A., Guo X., Mendez R., Parsaud H, & Hickman J.J. (2023). The Effect of Skeletal Muscle-Specific Creatine Treatment on ALS NMJ Integrity and Function. International Journal of Molecular Sciences, 24(17), 13519.

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Mechanical Loading Effects on NG2/CSPG4 Fibrochondrocytes

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To define the role of mechanical loading on NG2/CSPG4 functionality, primary mandibular fibrochondrocytes were seeded in agarose-collagen scaffold and loaded in a compression bioreactor as previously described (15 (link)). In short, cell-agarose-collagen scaffolds were generated from a 5% low-gelling temperature agarose (A0701, Sigma, Burlington, MA) mixed in 5 ml 1× PBS. Once cooled to 45 °C, the 5% agarose solution was combined with 0.25 mg/ml rat tail collagen (A1048301, Thermo Fisher, Waltham, MA) and a cell suspension in 1 ml FBS supplemented DMEM to create a 4% agarose/collagen solution. A cell density of 4 × 10⁵/mm³ was used for all experiments (50 (link)). For casting, the cell-agarose/collagen solution was poured on a 1 mm glass spacer plate and covered to generate a uniform 1 mm thickness sheet (1653311, Bio-Rad, Des Plaines, IL). A punch was used to generate plugs measuring 17 mm in radius. The cell-agarose/collagen scaffold plugs cultured for 72 h in supplemented advanced DMEM (12492-013, Gibco, Gaithersburg, MD) at 37 °C and 5% CO₂. For loading, all cell-agarose-collagen scaffolds were put in a compression bioreactor housed inside of a cell incubator and loaded in constrained, uniaxial compression at 2.5 N for 2 h. All loaded samples were compared to unloaded controls.

Reed D.A., Zhao Y., Bagheri Varzaneh M., Shin J.S., Rozynek J., Miloro M, & Han M. (2022). NG2/CSPG4 regulates cartilage degeneration during TMJ osteoarthritis. Frontiers in dental medicine, 3, 1004942.

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Synthesis of Functionalized Graphene Oxide-Collagen Composites

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A 4 mg/mL GO aqueous solution (Sigma-Aldrich, 777676) was first diluted with deionized water to 300 μg/mL and placed in an oven at 80 °C for 3, 5, and 9 days. The solution was then dispersed using an ultrasonic probe sonicator and filtered using a 1.2 μm syringe filter to obtain the aGO solution. For the synthesis of aGO-COL composites, carboxyl groups of the aGO solution were activated through the EDC/NHS coupling reaction for 1 h; it was then mixed with type I collagen from rat tails (Thermo Fisher Scientific, A1048301) for 1 h. Three different aGO weight percentages in aGO-COL were used in the experiments: 16%, 32%, and 64% (w/w, aGO/COL).

Yang J.W., Chen C.Y., Yu Z.Y., Chung J.H., Liu X., Wu C.Y, & Chen G.Y. (2022). An electroactive hybrid biointerface for enhancing neuronal differentiation and axonal outgrowth on bio-subretinal chip. Materials Today Bio, 14, 100253.

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Collagen Monolayer Preparation for MUSI-tAF Analysis

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To compare MUSI-tAF results with SEM, we required a monolayer of collagen. We used rat tail collagen-I for this purpose (A1048301, Thermo Fisher Scientific). The collagen was mixed with 10

\times

PBS and sodium hydroxide as per manufacturers’ instructions to prepare a final concentration of 0.2 mg/ml. 200

μ

l of the collagen solution was allowed to dry in coverslips for 24 hours. After the drying stage, the collagen was washed 5

\times

in distilled water to remove any excess salt deposits that may interfere with imaging. The collagen was allowed to air dry at room temperature for 48 hours before imaging. Ideally, the sample is a monolayer but we expect several areas to be 3-5 collagen fibril thickness.

Ghosh B., Chatterjee J., Paul R.R., Acuña S., Lahiri P., Pal M., Mitra P, & Agarwal K. (2024). Molecular histopathology of matrix proteins through autofluorescence super-resolution microscopy. Scientific Reports, 14, 10524.

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Collagen-Induced Platelet Aggregation Assay

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The rats were anaesthetised with sodium thiopental (100 mg/kg, i.p.). Collagen solution (collagen I, rat tail, A1048301, Thermo Fisher Scientific, Inc.) was diluted with physiological saline (saline, Otsuka Pharmaceutical Factory, Inc.) at concentrations of 0.1, 0.3, and 1 mg/mL. The saline served as a vehicle. The collagen solutions (1 mL/kg) or their vehicle were administered to the rats intravenously as a bolus to the jugular vein. Blood was collected from the jugular vein into a syringe containing 10 vol% of 3.8% sodium citrate before and 10, 30, 60, 120, and 180 min after collagen injection, and platelets were counted with an automated haematology analyser (Cellutac α, Nihon Kohden Corporation). The change in platelet count was expressed as the percentage change from before collagen injection, and the AUC of platelet count (% · min) was also calculated.

Ito Y., Ohno K., Morikawa Y., Tomizawa A., Mizuno M, & Sugidachi A. (2018). Vasodilator-stimulated phosphoprotein (VASP) is not a major mediator of platelet aggregation, thrombogenesis, haemostasis, and antiplatelet effect of prasugrel in rats. Scientific Reports, 8, 9955.

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ADCY9 and BVES Regulation of cAMP Signaling

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The ADCY3/ADCY6 double KO HEK293 cells with stable expression of cAMPr were seeded in a 35-mm glass-bottom dish coated with collagen (A1048301, Thermo Fisher Scientific, Waltham, MA). The cells were transfected with the indicated vectors including pmCherry-C1, pCMV-ADCY9-Flag and pCMV-BVES-HA. At 24 h after transfection, the cells were washed twice with 1x Hanks’ balanced salt solution and imaged under a Nikon Ti-E fluorescence microscope (Nikon, Melville, NY), equipped with a Zyla 5.5 sCMOS camera (Andor, Concord, MA) as well as NIS-Elements AR version 4.50 (Nikon, Melville, NY). Time series images were acquired every 4 s for 8 min. 500 μM IBMX was added to the culture dish at about 30 s after the fluorescence signal reaching a stable baseline. The cAMPr fluorescence signal data after background subtraction were plotted using Graphpad Prism 8.0.1.

Li H., Wang P., Zhang C., Zuo Y., Zhou Y, & Han R. (2023). Defective BVES-mediated feedback control of cAMP in muscular dystrophy. Nature Communications, 14, 1785.

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Fabrication and Preparation of NMJ Chambers

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Polydimethylsilxane (PDMS) chambers were designed, casted and cut as previously described.^{[21a (link)]} The PDMS NMJ chambers were incubated for 24 hours in 70% isopropanol to remove any unpolymerized monomers that could be toxic to the cells. The chambers were dipped in 100% ethanol and dried under sterile conditions for at least 2-hours prior to assembly. Glass coverslips (22 mm x 22 mm) were cleaned by plasma treatment for 2-minutes under oxygen pressure of 750 mTorr, sterilized with 70% ethanol and air-dried under sterile conditions before assembly. The sterile PDMS chambers adhered to the clean coverslips by applying gentle pressure around the edges and gently tapping on the tunnels. The SKM-side and MN-side of the chamber were coated with rat tail collagen I (ThermoFisher A1048301; 60 μg/mL) and laminin (ThermoFisher 23017015; 3 μg/mL), respectively. The collagen was removed after a 2-hour incubation period and rinsed twice with 1X phosphate buffer solution (PBS). The muscle proliferation medium (Adult Growth Medium, AGM) was added to the muscle-side before storing the systems at 4°C overnight. The laminin remained on the MN-side. After 24 hours, the laminin solution on the MN-side was removed and replaced with human motoneuron media (HMN). The NMJ systems were stored at 37°C and 5% CO₂ 1-hour before cell culture plating for equilibration.

Guo X., Smith V., Jackson M., Tran M., Thomas M., Patel A., Lorusso E., Nimbalkar S., Cai Y., McAleer C.W., Wang Y., Long C.J, & Hickman J.J. (2020). A Human-Based Functional NMJ System for Personalized ALS Modeling and Drug Testing. Advanced therapeutics, 3(11), 2000133.

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