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125i aβ40

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125I-Aβ40 is a radioactive tracer used for research purposes. It is composed of the amyloid-beta 40 peptide labeled with the radioactive isotope iodine-125. This product is intended for use in in vitro research applications.

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Lab products found in correlation

14c inulin, by American Radiolabeled Chemicals (4 mentions) 14c inulin, by PerkinElmer (3 mentions) Lectin, by Vector Laboratories (1 mentions) Wallac 1414 winspectral counter, by PerkinElmer (1 mentions) Wallac 1470 wizard gamma counter, by PerkinElmer (1 mentions) Valspodar, by Xenotech (1 mentions) Ketamine, by Henry Schein (1 mentions) Xylazine, by Henry Schein (1 mentions)

5 protocols using 125i aβ40

1

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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2

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 (BCItotal(%)) and clearance of 125I-Aβ40 across
BBB (BCIBBB(%)) 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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3

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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4

Measurement of Aβ40 Clearance in Mice

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In vivo Aβ40 clearance was investigated using the BEI method in C57BL/6 wild-type mice as described previously.53 (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 (link) (30 nM, PerkinElmer, MA) and 14C-inulin (0.02 mCi, American Radiolabeled Chemicals, St. Louis, MO) prepared in extracellular fluid buffer (ECF) was microinjected in the mice brains. Thirty minutes later, brains were rapidly collected for 125I-Aβ40 analysis and microvessels isolation as described below. Calculations of 125I-Aβ40 clearance were performed as described previously.53 (link) 125I-Aβ40 and 14C-inulin radioactivity were determined in brain tissues using a Wallac beta and gamma counter. 125I-Aβ40 BEI% was determined as described previously.53 (link)
Batarseh Y.S., Bharate S.S., Kumar V., Kumar A., Vishwakarma R.A., Bharate S.B, & Kaddoumi A. (2017). Crocus sativus Extract Tightens the Blood-Brain Barrier, Reduces Amyloid β Load and Related Toxicity in 5XFAD Mice. ACS chemical neuroscience, 8(8), 1756-1766.
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5

Investigating Aβ40 Clearance In Vivo

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In vivo Aβ40 clearance was investigated using the BEI method as described previously (Qosa et al., 2012 (link)). In brief, a stainless steel guide cannula was implanted stereotaxically into the right caudate nucleus of mice that had been anesthetized with intraperitoneal xylazine and ketamine (20 and 125 mg/kg, respectively) (Henry Schein Inc., NY). After 12 h recovery period, animals were re-anesthetized and tracer fluid (0.5 μl) containing 125I-Aβ40 (30 nM, PerkinElmer, MA) and 14C-inulin (0.02 μCi, American Radiolabeled Chemicals, MO) prepared in extracellular fluid buffer (ECF) was administered. Thirty minutes post 125I-Aβ40 injection (Cirrito et al., 2005 (link); Shibata et al., 2000 (link)); brain tissues were rapidly collected for 125I-Aβ40 analysis. To characterize role of P-gp and LRP1, 0.5 μl of ECF containing valspodar (40 μM; XenoTech, KS), a well-established P-gp inhibitor, or RAP (1 μM; Oxford Biomedical Research, MI), an LRP1 inhibitor, were intracerebrally administered 5 min prior to 125I-Aβ40 injection.
Qosa H., Abuasal B.S., Romero I.A., Weksler B., Couraud P.O., Keller J.N, & Kaddoumi A. (2014). Differences in amyloid-β clearance across mouse and human blood-brain barrier models: Kinetic analysis and mechanistic modeling. Neuropharmacology, 79, 668-678.
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