Hen egg ovalbumin

Manufactured by GE Healthcare

Hen egg ovalbumin is a naturally occurring protein derived from chicken eggs. It serves as a standard reference material for various analytical and research applications.

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Lab products found in correlation

Sephadex g 25, by GE Healthcare (2 mentions) Alexa fluor 488 c5 maleimide, by Thermo Fisher Scientific (2 mentions) Superdex 200 10 300 column, by GE Healthcare (1 mentions) Superdex 200 10 300 gl column, by GE Healthcare (1 mentions) Yeast alcohol dehydrogenase, by Merck Group (1 mentions)

3 protocols using hen egg ovalbumin

Fluorescent Labeling of PlxnD1 Protein

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PlxnD1 Y517C, A1135C at a concentration of 10 µM in PBS was labelled with a 20-fold molar excess of a thiol-reactive fluorescent dye, Alexa Fluor 488 C5 maleimide (ThermoFisher), and the reaction was allowed to proceed for 24 h at 6°C in the dark. Unreacted dye was removed from the labelled protein using a Sephadex G-25 (GE Healthcare). The degree of labelling (n) was determined using the following formula (Eq.1):

n = \frac{A_{488} M_{w}}{ε c}

where A₄₈₈ is the absorbance at 488 nm, M_w is the molecular weight of protein, ε is the molar extinction coefficient of the dye and c is the protein concentration in mg/ml. Hen egg ovalbumin (GE Healthcare) was used as a positive control.

Mehta V., Pang K.L., Rozbesky D., Nather K., Keen A., Lachowski D., Kong Y., Karia D., Ameismeier M., Huang J., Fang Y., del Rio Hernandez A., Reader J.S., Yvonne Jones E, & Tzima E. (2020). The Guidance Receptor Plexin D1 Moonlights as an Endothelial Mechanosensor. Nature, 578(7794), 290-295.

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Fluorescent Labeling of PlxnD1 Protein

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n = \frac{A_{488} M_{w}}{ε c}

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Analytical SEC of KtrC with Ligands

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Analytical size exclusion chromatography was performed using a Superdex 200 10/300 GL column (GE Healthcare). For each analysis, 500 μl of KtrC at 10, 6 and 3 mg/ml in the presence of 5 mM ATP or ADP and 5 mM MgCl2 were loaded into the column equilibrated with 50 mM Tris-HCl pH 8.0, 150 mM KCl, 5 mM DTT, 0.5 mM MgCl2, at a flow rate of 0.5 ml/min. Size-exclusion runs were also performed in the absence of MgCl2 either in the protein sample or elution buffer. Bacillus subtilis KtrA (199 kDa), yeast alcohol dehydrogenase (147 kDa; Sigma), chicken egg white Conalbumin (75 kDa) and hen egg Ovalbumin (43 kDa; both from GE Healthcare) were used as molecular weight standards for gel filtration calibration KtrC-ATP-Mg 2+ and KtrC-ADP-Mg 2+ (8 mg/ml) were incubated with 0.2 mM c-di-AMP. Complex formation was analyzed by size exclusion chromatography. For this, 50 L of protein sample (400 µg) were injected in a Superdex 200 10/300 column (GE Healthcare) equilibrated with 50 mM Tris-HCl pH 8.0, 150 mM KCl, 5 mM DTT.

Rocha R., Teixeira-Duarte C.M., Jorge J.M.P, & Morais-Cabral J.H. (2019). Characterization of the molecular properties of KtrC, a second RCK domain that regulates a Ktr channel in Bacillus subtilis. Journal of structural biology, 205(3).

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