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ea002 kit  (R&D Systems)


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    R&D Systems ea002 kit
    Ea002 Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sialyltransferase+activity+kit/Sialyltransferase+Activity+Kit/pm40324300-624-35-37
    Average 93 stars, based on 14 article reviews
    ea002 kit - by Bioz Stars, 2026-10
    93/100 stars

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    Related Articles

    Activity Assay:

    Article Title: Transfer of Functional Cargo in Exomeres
    Article Snippet: Protein concentration was quantified using BCA (Thermo Fischer Scientific/Pierce, Waltham, MA). .. Sialyltransferase activity present in the lysates was determined using the Sialyltransferase Activity Kit from R&D Systems (cat# EA002) according to the vendor protocol with a few modifications Briefly, CMP-Sialic Acid (Sigma, cat# C8271) was used at a final concentration of 2.5 mM, N -acetyl-D-lactosamine (Sigma, cat# A7791) was used at a final concentration of 2.5 mM, the coupling phosphatase was used at a final concentration of 10 ng/μL, and CMP was used at a final concentration of 0.1 mM. ..

    Article Title: Engineering of CHO cells for the production of vertebrate recombinant sialyltransferases
    Article Snippet: Images were acquired using an upright microscope (Olympus BX53) with violet (autofluorescence/DAPI) and green (FITC) filters. .. The Sialyltransferase Activity Kit (R&D Systems, Abingdon, UK) was used essentially as described by the manufacturer and as described elsewhere ( ; ). ..

    Article Title: A universal glycoenzyme biosynthesis pipeline that enables efficient cell-free remodeling of glycans
    Article Snippet: Imaging of blots was performed using a ChemiDoc TM XRS + System following a brief incubation with Western ECL substrate (Bio-Rad). .. Kinetic analysis of sialyltransferases was performed using a commercial sialyltransferase activity kit (R&D Systems, Cat # EA002) according to the manufacturer’s protocols. .. Briefly, assays used 2 μg/mL of purified Sx-Δ26 Hs ST6Gal1 or commercial human ST6Gal1 (amino acids 44–406) (R&D Systems; Cat # 7620-GT-010), 1.0 mg/mL of asialofetuin (Sigma-Aldrich; Cat # A4781-50MG) as acceptor substrate, and 0.02–0.8 mM of CMP-Neu5Ac as donor substrate.

    Article Title: Effects of autophagy-inhibiting chemicals on sialylation of Fc-fusion glycoprotein in recombinant CHO cells.
    Article Snippet: .. The enzyme activities of intracellular/extracellular sialidase, as well as intracellular α2,3-sialyltransferase (α2,3-ST), were measured by the Amplex Red Neuraminidase Assay Kit (Thermo Fisher Scientific) and the Sialyltransferase Activity Kit (R&D Systems, MN, USA), respectively, following the manufacturer’s instructions. ..

    Article Title: The androgen receptor controls expression of the cancer-associated sTn antigen and cell adhesion through induction of ST6GalNAc1 in prostate cancer
    Article Snippet: FLAG-tagged ST6GalNAc1 full length and ST6GalNAc1-short were immunoprecipitated from Flp-InTM-293 cell lysates using EZview Red ANTI-FLAG M2 Affinity gel (Sigma F2426). .. Sialyltransferase assays were performed using the Sialyltransferase Activity Kit (R&D Systems EA002) according to the manufacturer's instructions using 25nmol of CMP-Neu5Ac (Sigma), 0.5mg asialofetuin (Sigma) and 50ng of Coupling Phosphatise 2 for 20 minutes at 37°C. ..

    Article Title: A universal glycoenzyme biosynthesis pipeline that enables efficient cell-free remodeling of glycans.
    Article Snippet: Imaging of blots was performed using a ChemiDocTM XRS+ System following a brief incubation with Western ECL substrate (Bio-Rad). .. Kinetic analysis of sialyltransferases was performed using a commercial sialyltransferase activity kit (R&D Systems, Cat # EA002) according to the manufacturer’s protocols. .. Briefly, assays used 2μg/mL of purified Sx-Δ26HsST6Gal1 or commercial human ST6Gal1 (amino acids 44–406) (R&D Systems; Cat # 7620-GT-010), 1.0mg/mL of asialofetuin (Sigma-Aldrich; Cat # A4781-50MG) as acceptor substrate, and 0.02–0.8mM of CMP-Neu5Ac as donor substrate.

    Article Title: Effects of autophagy-inhibiting chemicals on sialylation of Fc-fusion glycoprotein in recombinant CHO cells
    Article Snippet: .. The enzyme activities of intracellular/extracellular sialidase, as well as intracellular α2,3-sialyltransferase (α2,3-ST), were measured by the Amplex Red Neuraminidase Assay Kit (Thermo Fisher Scientific) and the Sialyltransferase Activity Kit (R&D Systems, MN, USA), respectively, following the manufacturer’s instructions. ..

    Article Title: Utilization of Glucocorticoids as Additives for Enhanced Sialylation of Fc-fusion Protein in CHO Cell Cultures
    Article Snippet: Sialylation, which is considered as a critical quality attribute, plays important roles in pharmacokinetics of recombinant proteins.. Terminal sialic acids of glycoproteins are modulated by intracellular sialyltransferase (ST) and extracellular sialidase.. Since CHO cells producing recombinant proteins can only transfer α2,3-sialic acid to a nascent oligosaccharide, inactivation of α2,6-ST results in a different composition of glycan with human proteins, which have predominantly α2,6-linkages.

    Concentration Assay:

    Article Title: Transfer of Functional Cargo in Exomeres
    Article Snippet: Protein concentration was quantified using BCA (Thermo Fischer Scientific/Pierce, Waltham, MA). .. Sialyltransferase activity present in the lysates was determined using the Sialyltransferase Activity Kit from R&D Systems (cat# EA002) according to the vendor protocol with a few modifications Briefly, CMP-Sialic Acid (Sigma, cat# C8271) was used at a final concentration of 2.5 mM, N -acetyl-D-lactosamine (Sigma, cat# A7791) was used at a final concentration of 2.5 mM, the coupling phosphatase was used at a final concentration of 10 ng/μL, and CMP was used at a final concentration of 0.1 mM. ..



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    A malachite-green-based <t>nucleotidase-coupled</t> assay measures the activity of purified Cho1 protein. ( A ) Blue native PAGE gel of the purified hexameric tag-free Cho1 protein. Purified Cho1 and protein ladder with known MW are indicated. The gel was stained with Coomassie Blue R-250. ( B ) Schematic representation of the malachite-green-based nucleotidase-couple assay. Cho1 synthesizes PS from CDP-DAG (cytidyldiphosphate-diacylglycerol) and serine. This releases PS and CMP (cytidylmonophosphate). The phosphate from CMP is cleaved by the nucleotidase <t>CD73</t> to release inorganic phosphate, which can be bound by the malachite green reagent and measured colorimetrically at OD 620 . AB-680 is a potent inhibitor of CD73 and can, thus, inhibit the reaction. ( C ) OD 620 signal from the malachite green reagent that was added to the reaction (shown in B ) at different time points after the reaction started. Reactions were set up with the same conditions and stopped by adding malachite green at the time indicated. The dots represent the mean of four biological replicates, and the error bars are ±standard deviation (S.D.) values. ( D ) Inhibition of the nucleotidase-coupled assay by AB-680 is shown for a series of replicates in 384-well format and ( E ) is quantified for a total of 21 replicates. Statistics were conducted using one-way ANOVA using Tukey’s multiple comparisons test (ns, not significant; **** P < 0.0001).
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    Image Search Results


    A malachite-green-based nucleotidase-coupled assay measures the activity of purified Cho1 protein. ( A ) Blue native PAGE gel of the purified hexameric tag-free Cho1 protein. Purified Cho1 and protein ladder with known MW are indicated. The gel was stained with Coomassie Blue R-250. ( B ) Schematic representation of the malachite-green-based nucleotidase-couple assay. Cho1 synthesizes PS from CDP-DAG (cytidyldiphosphate-diacylglycerol) and serine. This releases PS and CMP (cytidylmonophosphate). The phosphate from CMP is cleaved by the nucleotidase CD73 to release inorganic phosphate, which can be bound by the malachite green reagent and measured colorimetrically at OD 620 . AB-680 is a potent inhibitor of CD73 and can, thus, inhibit the reaction. ( C ) OD 620 signal from the malachite green reagent that was added to the reaction (shown in B ) at different time points after the reaction started. Reactions were set up with the same conditions and stopped by adding malachite green at the time indicated. The dots represent the mean of four biological replicates, and the error bars are ±standard deviation (S.D.) values. ( D ) Inhibition of the nucleotidase-coupled assay by AB-680 is shown for a series of replicates in 384-well format and ( E ) is quantified for a total of 21 replicates. Statistics were conducted using one-way ANOVA using Tukey’s multiple comparisons test (ns, not significant; **** P < 0.0001).

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    Figure Lengend Snippet: A malachite-green-based nucleotidase-coupled assay measures the activity of purified Cho1 protein. ( A ) Blue native PAGE gel of the purified hexameric tag-free Cho1 protein. Purified Cho1 and protein ladder with known MW are indicated. The gel was stained with Coomassie Blue R-250. ( B ) Schematic representation of the malachite-green-based nucleotidase-couple assay. Cho1 synthesizes PS from CDP-DAG (cytidyldiphosphate-diacylglycerol) and serine. This releases PS and CMP (cytidylmonophosphate). The phosphate from CMP is cleaved by the nucleotidase CD73 to release inorganic phosphate, which can be bound by the malachite green reagent and measured colorimetrically at OD 620 . AB-680 is a potent inhibitor of CD73 and can, thus, inhibit the reaction. ( C ) OD 620 signal from the malachite green reagent that was added to the reaction (shown in B ) at different time points after the reaction started. Reactions were set up with the same conditions and stopped by adding malachite green at the time indicated. The dots represent the mean of four biological replicates, and the error bars are ±standard deviation (S.D.) values. ( D ) Inhibition of the nucleotidase-coupled assay by AB-680 is shown for a series of replicates in 384-well format and ( E ) is quantified for a total of 21 replicates. Statistics were conducted using one-way ANOVA using Tukey’s multiple comparisons test (ns, not significant; **** P < 0.0001).

    Article Snippet: A total of 30–35 ng of purified Cho1 protein was used in each reaction in 50 mM Tris-HCl (pH 8.0) for primary screening, combined with 100 μM CDP-DAG (Avanti, cat# 870510), 5 mM serine, 0.4 ng CD73 nucleotidase (R&D systems, cat# EA002), 1 mM MnCl 2 , 0.1% APX-100, and 0.1% digitonin, in a total volume of 10 μL.

    Techniques: Activity Assay, Purification, Blue Native PAGE, Staining, Standard Deviation, Inhibition