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Calibration standards

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Calibration standards are reference materials used to verify and validate the accuracy and precision of laboratory equipment. They provide known quantities or concentrations of specific analytes, allowing for the calibration and performance verification of instruments and test methods.

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Lab products found in correlation

Flag peptide, by Merck Group (2 mentions) Ezview red anti flag m2 affinity gel, by Merck Group (2 mentions) Superdex 75 hr 10 30 column, by GE Healthcare (2 mentions)

Spelling variants of this product

Gel filtration calibration standards (2 citations)

2 protocols using calibration standards

1

Isolation and characterization of FLAG-CtIP complexes

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Nuclear extracts from HEK-293T cells transiently transfected with plasmids expressing wild type or L27E versions of FLAG-CtIP were prepared as described previously48 (link). NaCl concentration in extracts was adjusted to 300 mM and FLAG-tagged proteins were isolated by EZview Red anti-FLAG M2 Affinity gel (Sigma). Anti-FLAG beads were washed three times with lysis buffer containing 300 mM NaCl. Following elution with 3×FLAG peptide (Sigma) proteins were fractionated on a Superdex 75 HR 10/30 column (GE Healthcare). Relative molecular weights of the FLAG-CtIP protein complexes were calculated based on the elution profiles of the calibration standards (GE Healthcare), which were as follows: thyroglobulin (669 kDa), ferritin (440 kDa), aldolase (158 kDa), conalbumin (75 kDa), ovalbumin (44 kDa).
Davies O.R., Forment J.V., Sun M., Belotserkovskaya R., Coates J., Galanty Y., Demir M., Morton C.R., Rzechorzek N.J., Jackson S.P, & Pellegrini L. (2015). CtIP tetramer assembly is required for DNA-end resection and repair. Nature structural & molecular biology, 22(2), 150-157.
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2

Isolation and characterization of FLAG-CtIP complexes

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Nuclear extracts from HEK-293T cells transiently transfected with plasmids expressing wild type or L27E versions of FLAG-CtIP were prepared as described previously48 (link). NaCl concentration in extracts was adjusted to 300 mM and FLAG-tagged proteins were isolated by EZview Red anti-FLAG M2 Affinity gel (Sigma). Anti-FLAG beads were washed three times with lysis buffer containing 300 mM NaCl. Following elution with 3×FLAG peptide (Sigma) proteins were fractionated on a Superdex 75 HR 10/30 column (GE Healthcare). Relative molecular weights of the FLAG-CtIP protein complexes were calculated based on the elution profiles of the calibration standards (GE Healthcare), which were as follows: thyroglobulin (669 kDa), ferritin (440 kDa), aldolase (158 kDa), conalbumin (75 kDa), ovalbumin (44 kDa).
Davies O.R., Forment J.V., Sun M., Belotserkovskaya R., Coates J., Galanty Y., Demir M., Morton C.R., Rzechorzek N.J., Jackson S.P, & Pellegrini L. (2015). CtIP tetramer assembly is required for DNA-end resection and repair. Nature structural & molecular biology, 22(2), 150-157.
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