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Asparaginase

Manufactured by Abcam
Sourced in United States

Asparaginase is an enzyme that catalyzes the conversion of asparagine to aspartic acid and ammonia. It is commonly used in research applications.

Automatically generated - may contain errors

3 protocols using asparaginase

1

Fluorescent Dye Labeling Protocol

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All fluorescent dyes were purchased from ThermoFisher Scientific (Invitrogen, UK), and CPA was from Merck Millipore (Calbiochem, UK). Collagenase was obtained from Worthington (USA), asparaginase was from Abcam (UK), the PAR2 inhibitor FSLLRY-NH2 from TOCRIS (UK) and GSK-7975A from GlaxsoSmithKline (UK). All other chemicals were purchased from Sigma. C57BL/6 J mice were from Charles River UK Ltd.
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2

Asparaginase Treatment in Nude Mice

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The animal work was approved by the Institutional Animal Care and Use Committee (IACUC) at the Broad Institute. 4-week-old, female, athymic nude mice (CrTac:NCr-Foxn1nu, Taconic) were inoculated subcutaneously with 7*106 cancer cells in phenol red free RPMI media with 50% matrigel in both flanks. The mice were randomized into treatment or control group when tumors reached approximately 100–200 mm3 in size. Asparaginase (Abcam) was delivered with intraperitoneal injection at 3000 units/kg in 200 μl PBS 5 times per week (omitting Wednesday and Sunday) for 3 weeks. Tumor tissues were collected and processed for IHC staining by standard methods. All IHC staining was performed on the Leica Bond automated staining platform. Polyclonal Asparagine Synthetase (ASNS) antibody from Proteintech (#14861–1-AP) was run at 1:1500 dilution using the Leica Biosystems Refine Detection Kit with citrate antigen retrieval. Tumor sizes were calculated by 1/2*length*width*width.
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3

Fluorescent Dyes for Cell Analysis

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Fluorescent dyes Fluo-4-AM, MgGreen AM, and propidium iodide (PI) were purchased from Thermo Fisher Scientific. Collagenase was obtained from Worthington, asparaginase was purchased from Abcam, and POAEE was from Cayman Chemical. All other reagents were purchased from Sigma-Aldrich. C57BL/6J mice were obtained from The Jackson Laboratory.
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