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Artificial csf

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Artificial CSF is a sterile, buffered saline solution that mimics the composition of human cerebrospinal fluid (CSF). It is designed to maintain the physiological environment of cells and tissues typically found in the central nervous system.

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Lab products found in correlation

Alexa fluor 647, by Thermo Fisher Scientific (2 mentions) 40kd mwco zeba spin desalting columns, by Thermo Fisher Scientific (2 mentions) Mopc 21, by BioXCell (2 mentions) Formic acid, by Thermo Fisher Scientific (1 mentions) Ceftazidime pentahydrate, by Thermo Fisher Scientific (1 mentions) Methanol, by Thermo Fisher Scientific (1 mentions) 5 ipf2α 6 d11, by Cayman Chemical (1 mentions) 8 iso 15 r pgf2α, by Cayman Chemical (1 mentions) 15 r pgf2α, by Cayman Chemical (1 mentions) Aβ1 40, by Peptide Institute (1 mentions) Aβ1 42, by Peptide Institute (1 mentions) Milli q integral 10, by Merck Group (1 mentions)

5 protocols using artificial csf

1

Cefepime Quantification in Biological Fluids

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Animals were administered cefepime hydrochloride (Hospira, Lake Forest, IL [lot no. 144G005]) for injection. Artificial CSF (Tocris Bioscience, Bristol, UK) and normal saline (Abbott [lot no. C950212]) were used as in the sampling methods described below. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) standard curves were generated using commercially obtained cefepime hydrochloride (USP) (Chem-Impex International, Inc., Wood Dale, IL) with a purity of greater than 99.5%. Ceftazidime pentahydrate (Acros Organics, NJ) was used as an internal standard for the cefepime quantification. Formic acid was obtained from Thermo Fisher Scientific (Waltham, MA). Acetonitrile (BDH [lot no. 16I011254]) and methanol (BDH [lot no. 16L204024]) were obtained from VWR Analytical (Chicago, IL). Milli-Q water was obtained from Midwestern University via an Aqua Solutions water purification dispensatory. Frozen, nonmedicated, nonimmunized, pooled Sprague-Dawley rat plasma (anticoagulated with disodium EDTA) and CSF were used for calibration of standard curves (BioreclamationIVT, Westbury, NY).
Avedissian S.N., Pais G., Joshi M.D., Rhodes N.J, & Scheetz M.H. (2019). A Translational Pharmacokinetic Rat Model of Cerebral Spinal Fluid and Plasma Concentrations of Cefepime. mSphere, 4(1), e00595-18.
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2

Eicosanoid Standards Protocol

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Standards of F2-IP (5[R]-iPF2α-VI, 5[S]-iPF2α-VI, 5-iPF2α-VI-d11, 8-iso-15[S]-PGF2α, 8-iso-15[R]-PGF2α, 8-iso-15[S]-PGF2α-d4, and 15[R]-PGF2α), 15(S)-PGF2α (hereafter referred to as PGF2α), 20:4, and 20:4-d8 were from Cayman Chemicals (Ann Arbor, MI). Artificial CSF was from Tocris Bioscience (Bristol, UK). Methanol (Fisher Scientific Inc.), hexane, and ethyl acetate (Lab Scan) were of high-performance liquid chromatography grade, and Bond Elut CertifyII SPE columns were from Agilent Technologies (Santa Clara, CA). Other chemicals were from Sigma, unless indicated otherwise.
Lam M.A., Maghzal G.J., Khademi M., Piehl F., Ratzer R., Romme Christensen J., Sellebjerg F.T., Olsson T, & Stocker R. (2016). Absence of systemic oxidative stress and increased CSF prostaglandin F2α in progressive MS. Neurology® Neuroimmunology & Neuroinflammation, 3(4), e256.
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3

Intracerebral Delivery of Anti-CD22 Antibody

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A monoclonal anti-CD22 antibody (Cy34, BioXCell, 1mg/mL) or mouse IgG1 isotype control antibody (MOPC21, BioXCell, 1mg/mL) were conjugated to AlexaFluor647 (ThermoFisher) using NHS chemistry at a dye:protein molar ratio of 4. Free dye was removed and buffer was exchanged with artificial CSF (Tocris) using 40kD MWCO Zeba spin desalting columns (ThermoFisher). The labeled antibodies were loaded into 200 microliter osmotic pumps (Alzet/Durect) with a 28-day infusion rate of 0.25mL per hour. An infusion rate was chosen to maintain a steady state concentration of 10 μg/mL, a dose shown to promote phagocytosis with this particular antibody ex vivo. Osmotic pumps were connected to a cannula (Brain infusion kit III, Alzet) inserted at +1mm lateral, −0.3mm anterior-posterior, and −3mm deep relative to bregma in order to target the right lateral ventricle. The pump was placed subcutaneously and mice received post-surgical buprenorphine and Baytril.
Pluvinage J.V., Haney M.S., Smith B.A., Sun J., Iram T., Bonanno L., Li L., Lee D.P., Morgens D.W., Yang A.C., Shuken S.R., Gate D., Scott M., Khatri P., Luo J., Bertozzi C.R., Bassik M.C, & Wyss-Coray T. (2019). CD22 blockade restores homeostatic microglial phagocytosis in aging brains. Nature, 568(7751), 187-192.
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4

Intracerebral Delivery of Anti-CD22 Antibody

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A monoclonal anti-CD22 antibody (Cy34, BioXCell, 1mg/mL) or mouse IgG1 isotype control antibody (MOPC21, BioXCell, 1mg/mL) were conjugated to AlexaFluor647 (ThermoFisher) using NHS chemistry at a dye:protein molar ratio of 4. Free dye was removed and buffer was exchanged with artificial CSF (Tocris) using 40kD MWCO Zeba spin desalting columns (ThermoFisher). The labeled antibodies were loaded into 200 microliter osmotic pumps (Alzet/Durect) with a 28-day infusion rate of 0.25mL per hour. An infusion rate was chosen to maintain a steady state concentration of 10 μg/mL, a dose shown to promote phagocytosis with this particular antibody ex vivo. Osmotic pumps were connected to a cannula (Brain infusion kit III, Alzet) inserted at +1mm lateral, −0.3mm anterior-posterior, and −3mm deep relative to bregma in order to target the right lateral ventricle. The pump was placed subcutaneously and mice received post-surgical buprenorphine and Baytril.
Pluvinage J.V., Haney M.S., Smith B.A., Sun J., Iram T., Bonanno L., Li L., Lee D.P., Morgens D.W., Yang A.C., Shuken S.R., Gate D., Scott M., Khatri P., Luo J., Bertozzi C.R., Bassik M.C, & Wyss-Coray T. (2019). CD22 blockade restores homeostatic microglial phagocytosis in aging brains. Nature, 568(7751), 187-192.
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5

Synthetic Amyloid-Beta Peptide Quantification

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Synthetic human Aβ1-38, Aβ1-40, Aβ1-42, and Aβ1-43 were purchased from the Peptide Institute (Osaka, Japan). Uniformly stable isotope-labeled peptides manufactured by rPeptide (Watkinsville, GA, USA), corresponding to each peptide (Aβ1-38, Aβ1-40, Aβ1-42, and Aβ1-43), were used as internal standards (ISs). Artificial CSF was purchased from Tocris Bioscience (Bristol, UK), and rat plasma was purchased from Charles River Laboratories Japan (Yokohama, Japan). Other materials and reagents were purchased from Kanto Chemical (Tokyo, Japan), Kokusan Chemical (Tokyo, Japan), and Sigma-Aldrich (St. Louis, MO, USA). Ultrapure water was prepared using Milli-Q Integral 10 (Merck, Darmstadt, Germany).
Seino Y., Nakamura T., Harada T., Nakahata N., Kawarabayashi T., Ueda T., Takatama M, & Shoji M. (2021). Quantitative Measurement of Cerebrospinal Fluid Amyloid-β Species by Mass Spectrometry. Journal of Alzheimer's Disease, 79(2), 573-584.
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