14c inulin

Manufactured by PerkinElmer

Sourced in United States

14C-inulin is a radioactive tracer compound used in various scientific and medical applications. It is a form of the carbohydrate inulin that has been labeled with the radioactive carbon-14 isotope. 14C-inulin is commonly used for measuring glomerular filtration rate and other physiological processes.

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Lab products found in correlation

125i aβ40, by PerkinElmer (3 mentions) 14c inulin, by American Radiolabeled Chemicals (2 mentions) Wallac wizard2 2470, by PerkinElmer (1 mentions) Tri carb 2800tr liquid scintillation analyzer, by PerkinElmer (1 mentions) L ascorbic acid, by Merck Group (1 mentions) Lectin, by Vector Laboratories (1 mentions) Wallac 1414 winspectral counter, by PerkinElmer (1 mentions) Wallac 1470 wizard gamma counter, by PerkinElmer (1 mentions) Lovastatin, by Merck Group (1 mentions) Bifonazole, by Merck Group (1 mentions) Simvastatin, by Merck Group (1 mentions) Clofazimine, by Merck Group (1 mentions) 3h taurocholate, by PerkinElmer (1 mentions)

6 protocols using 14c inulin

In Vitro Aβ Transcytosis Assay

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In order to study Aβ transcytosis in vitro, a standard transport model was used [10 (link), 12 (link), 35 (link)]. [¹²⁵I]-Aβ1–42 (0.1 nM) (purchased from Phoenix Peptide) and 1-μCi/ml [¹⁴C] inulin (purchased from PerkinElmer), a marker for paracellular diffusion, were added to serum-free media supplemented with 550-nM hydrocortisone and 40-mM HEPES and incubated at 37 °C. To study brain-to-blood transport, 10- and 80-μl samples were taken from the luminal compartment 60 min after the addition of [¹²⁵I]-Aβ1–42 and [¹⁴C]-inulin to the abluminal compartment. To investigate the amount of intact [¹²⁵I]-Aβ1–42 transported to the luminal side, 80-μl 15% TCA was added to an 80-μl luminal media sample and incubated for 10 min at 4 °C. Samples were then centrifuged at 16,000 g for 10 min. Pellets (representing intact [¹²⁵I]-Aβ1–42) were counted for [¹²⁵I]. Probes were counted on a Wallac Wizard2 2470 automatic γ-counter (PerkinElmer) for [¹²⁵I] or on a Tri-Carb 2800 TR Liquid Scintillation Analyzer (PerkinElmer) for [¹⁴C]. Transport of intact [¹²⁵I]-Aβ1–42 across the monolayer was calculated as Aβ1–42 transcytosis quotient (TQ) using the following formula:

Aβ 1 - 42 TQ = ([^{125} I] - Aβ 1 - 42 luminal / [^{125} I] - Aβ 1 - 42 input) / ([^{14} C] - inulin acceptor / [^{14} C] - inulin input)

Van Gool B., Storck S.E., Reekmans S.M., Lechat B., Gordts P.L., Pradier L., Pietrzik C.U, & Roebroek A.J. (2019). LRP1 Has a Predominant Role in Production over Clearance of Aβ in a Mouse Model of Alzheimer’s Disease. Molecular Neurobiology, 56(10), 7234-7245.

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Preparation of Native Hemoglobin and Ascorbate

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A fresh stock of CFH was prepared before each experiment by dissolving 10 mg endotoxin-free native human hemoglobin (Cell Sciences, Canton, MA) in 10 ml sterile 0.9% NaCl. A fresh stock of ascorbate was prepared before each experiment by dissolving L-ascorbic acid (Sigma, St. Louis, MO) in sterile H₂O. EDTA was purchased from Mediatech/CellGro-Corning (Manassas, VA). ¹⁴C-inulin (MW ~ 5000–5500; 2 mCi/g) was purchased from Perkin Elmer Life Sciences (Waltham, WA).

Kuck J.L., Bastarache J.A., Shaver C.M., Fessel J.P., Dikalov S.I., May J.M, & Ware L.B. (2017). Ascorbic acid attenuates endothelial permeability triggered by cell-free hemoglobin. Biochemical and biophysical research communications, 495(1), 433-437.

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In-vivo Clearance of 125I-Aβ40 via BEI Method

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The in-vivo 125I-Aβ40 (PerkinElmer; Boston, MA, USA) clearance was investigated using the brain efflux index (BEI) method as we described previously [20 (link)]. In brief, at the end of the treatment, animals were intraperitoneally anesthetized with xylazine and ketamine (20 and 125 mg/kg, respectively), followed by the insertion of a stainless-steel guide cannula into the right caudate nucleus, an area rich with blood microvessels, of the mice’s brain following previously established protocols [58 (link)]. A tracer fluid (0.5 μL) containing 125I-Aβ40 (30 nM; PerkinElmer, MA, USA) and 14C-inulin (0.02 mCi, American Radiolabeled Chemicals, St. Louis, MO, USA), prepared in an artificial extracellular fluid buffer (ECF; 122 mM NaCl, 25 mM NaHCO3, 3 mM KCl, 1.4 mM CaCl2, 1.2 mM MgSO4, 0.4 mM K2HPO4,10 mM D-glucose, and 10 mM HEPES, pH 7.4), was microinjected in the mice brains. Thirty minutes later, the brains were rapidly collected for a 125I-Aβ40 analysis. 125I-Aβ40 and 14C-inulin radioactivity was determined in the brain tissues using Wallac gamma and beta counters (PerkinElmer Inc. Waltham, MA, USA), respectively. 125I-Aβ40 BEI was determined using the formula:

Duong Q.V., Kintzing M.L., Kintzing W.E., Abdallah I.M., Brannen A.D, & Kaddoumi A. (2019). Plasma Rich in Growth Factors (PRGF) Disrupt the Blood-Brain Barrier Integrity and Elevate Amyloid Pathology in the Brains of 5XFAD Mice. International Journal of Molecular Sciences, 20(6), 1489.

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Amyloid-Beta Labeling and Detection

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Aβ40-HiLyte Fluor 488 and Aβ42-HiLyte Fluor 488 were obtained from AnaSpec (Fremont, CA, USA), and prepared as a 0.05% solution in artificial CSF (aCSF) following the manufacturer’s instructions. Samples were diluted in aCSF, aliquoted, stored at −20°C and used within two days. HFIP (1, 1, 1, 3, 3, 3-hexafluoro-2-propanol) treated lyophilized Aβ40 and Aβ42 were also obtained from AnaSpec, dissolved in ammonium hydroxide (1%) at room temperature for 15 minutes with intermittent mixing, sonicated for 5 minutes, diluted in aCSF to 20 μM, aliquoted and stored at −80°C. Each aliquot was used immediately and only once. Cascade blue (CB) labeled dextran 10 kDa (fixable), fluorescein labeled dextran 40 kDa (fixable) and Texas Red labeled dextran 3 kDa (fixable) were obtained from Molecular Probe (Eugene, OR, USA) and dissolved in aCSF (2% solution). Lectin was obtained from Vector Laboratory (Lycopersicon esculentum (tomato); Burlingame CA, USA). ¹²⁵I-Aβ40 and ¹⁴C-inulin were obtained from PerkinElmer (Waltham, MA, USA). We used ¹²⁵I-Aβ40 without added aprotinin, a potential inhibitor of LRP1-mediated transport. ELISA kits for Aβ40 (KHB 3481) and Aβ42 (KHB 3441) were obtained from Invitrogen (Camarilla, CA, USA) and Aβ oligomer ELISA kit (BEK-2215-1P) from Biosensis (Termecula, CA, USA).

Peng W., Achariyar T.M., Li B., Liao Y., Mestre H., Hitomi E., Regan S., Kasper T., Peng S., Ding F., Benveniste H., Nedergaard M, & Deane R. (2016). Suppression of glymphatic fluid transport in a mouse model of Alzheimer’s disease. Neurobiology of disease, 93, 215-225.

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In Vivo Amyloid-beta Clearance Assay

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In vivo Aβ₄₀ clearance was investigated using the
BCI method as described previously.^{36 (link)} Animals
were anesthetized followed by the insertion of a stainless steel guide
cannula into the right caudate nucleus of mice brains. A tracer fluid
(0.5 μL) containing ¹²⁵I-Aβ₄₀ (30
nM, PerkinElmer, MA) and ¹⁴C-inulin (0.02 mCi, American
Radiolabeled Chemicals, MO) prepared in extracellular fluid buffer
(ECF) was microinjected. Thirty minutes later, brains were rapidly
collected. One hemisphere of the brain was used for ¹²⁵I-Aβ₄₀ analysis and the second hemisphere was used
for microvessels isolation as described below. Calculations of ¹²⁵I-Aβ₄₀ clearance were performed as described
previously.^{36 (link)} Using trichloroacetic acid
(TCA) precipitation intact (precipitate) and degraded (supernatant) ¹²⁵I-Aβ₄₀ were determined in brain tissue
using a Wallac 1470 Wizard Gamma Counter (PerkinElmer, MA). ¹⁴C-Inulin in the precipitate and supernatant were also determined
using a Wallac 1414 WinSpectral Counter (PerkinElmer). The ¹²⁵I-Aβ₄₀ brain clearance index (BCI_total(%)) and clearance of ¹²⁵I-Aβ₄₀ across
BBB (BCI_BBB(%)) were determined as described previously.^{36 (link)}

Qosa H., Batarseh Y.S., Mohyeldin M.M., El Sayed K.A., Keller J.N, & Kaddoumi A. (2015). Oleocanthal Enhances Amyloid-β Clearance from the Brains of TgSwDI Mice and in Vitro across a Human Blood-Brain Barrier Model. ACS Chemical Neuroscience, 6(11), 1849-1859.

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Prestwick Chemical Library Screening

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The Prestwick chemical library (1280 FDA approved compounds in 96-well plates) was purchased from Prestwick Chemical (Illkirch, France). Bifonazole, bromhexine HCl, clofazimine, lovastatin, meclozine 2HCL, and simvastatin were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands). The fibroblast growth factor (FGF)19 human enzyme-linked immunosorbent assay kit was purchased from BioVendor R&D (Brno, Czech Republic). [³H]-taurocholate and [¹⁴C]-inulin were purchased from Perkin Elmer (Groningen, The Netherlands).

van de Wiel S.M., de Waart D.R., Oude Elferink R.P, & van de Graaf S.F. (2017). Intestinal Farnesoid X Receptor Activation by Pharmacologic Inhibition of the Organic Solute Transporter α-β. Cellular and Molecular Gastroenterology and Hepatology, 5(3), 223-237.

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