Endosafe pts spectrophotometer

Manufactured by Charles River Laboratories

The Endosafe‐PTS spectrophotometer is a laboratory instrument designed to measure the absorbance of samples. It utilizes a spectrophotometric method to determine the concentration of specific substances within a sample.

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Lab products found in correlation

Nickel column, by GE Healthcare (1 mentions) Pqe30xa, by Qiagen (1 mentions)

2 protocols using endosafe pts spectrophotometer

Production and Purification of Recombinant Proteins

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The mouse perlecan fragment LG3 (aa 3514‐3707 with N‐terminal His₈G tag) or a secreted form of red fluorescent protein (RFP1) (with C‐terminal His₈G tag) were cloned into the pTT5™ plasmid.26, 27 For protein production, 293‐6E cells were transfected at 1.8x10⁶cells/ml with pTT5/cDNA constructs. Cultures were harvested at 5 days posttransfection and purified by IMAC on Fractogel®‐Cobalt.28 For some experiments, N‐terminal His8G tag was removed from the LG3 construct.
Purified mouse serum albumin (MSA) protein was obtained from Alpha Diagnostic International (San Antonio, TX). Endotoxin levels were measured by Limulus Amebocyte Lysate test using Endosafe‐PTS spectrophotometer (Charles River Laboratories, Wilmington, MA) for LG3 (with or without His8G tag) and RFP1 and by clot method for MSA. The levels of endotoxin were respectively equal to 0.016 EU/mg, 0.021 EU/mg, and 1.2 EU/mg.

Padet L., Dieudé M., Karakeussian‐Rimbaud A., Yang B., Turgeon J., Cailhier J., Cardinal H, & Hébert M. (2018). New insights into immune mechanisms of antiperlecan/LG3 antibody production: Importance of T cells and innate B1 cells. American Journal of Transplantation, 19(3), 699-712.

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Purification and Characterization of AnxA5 and M1234

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AnxA5 and M1234 were prepared as described previously[14 (link)]. Briefly, AnxA5 and M1234 were expressed in Escherichia coli M15 (pREP4) (Qiagen, Venlo, the Netherlands), which were transformed with pQE30Xa (Qiagen) containing cDNA of human AnxA5 and M1234. Bacteria were harvested and lysed by sonification. Cell debris was removed by centrifugation. His-tagged proteins were isolated from supernatant by chromatography using nickel columns (GE Healthcare, Eindhoven, the Netherlands) and an imidazole gradient. Purified proteins were checked on homogeneity (MALDI-TOF/TOF) and PS binding activity (ellipsometry) as described elsewhere. All recombinant protein samples contained less than one endotoxin unit per ml as determined by Endosafe PTS spectrophotometer (Charles River, Leiden, the Netherlands).

Stöhr R., Schurgers L., van Gorp R., Jaminon A., Marx N, & Reutelingsperger C. (2017). Annexin A5 reduces early plaque formation in ApoE -/- mice. PLoS ONE, 12(12), e0190229.

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