Gradient native page gel

Manufactured by Thermo Fisher Scientific

The 4–16% gradient native PAGE gel is a laboratory equipment used for the separation and analysis of proteins based on their native (non-denatured) structure and charge. It provides a range of pore sizes that can accommodate a variety of protein sizes and complexes. The gradient allows for efficient separation and resolution of protein samples.

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Lab products found in correlation

Odyssey infrared imaging system, by LI COR (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions)

Close spelling variants of this product

Native PAGE 4–20% gradient TBE gels Native PAGE bis-tris gels of gradient 4 to 16%T Gradient native gel 6% native PAGE PAGE gels 4–12% gradient gels

2 protocols using gradient native page gel

Mitochondrial Respiratory Chain Complex Activities Assay

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Mitochondrial respiratory chain complex activities were determined by clear native (CN) PAGE In-gel activity assays^{44 (link)}. Yeast mitochondria (30 µg of protein for complex II, 90 µg for complexes III and IV, and 20 µg for complex V) were solubilized for 15 min on ice in solubilization buffer (1xNative PAGE buffer and 1% digitonin). The solubilized fraction was resolved using 4–16% gradient native PAGE gel (Life Technologies) with the following additions to the cathode buffer: for complex II, 0.05% sodium deoxycholate (DOC) and 0.01% n-dodecyl β-d-maltoside (DDM); for complex III, no addition; for complex IV, 0.05% DOC and 0.05% DDM; and for complex V, 0.05% DOC and 0.01% DDM were added. The gels were incubated in their respective activity staining solutions. Equal loading was determined by Coomassie blue staining.

Zulkifli M., Neff J.K., Timbalia S.A., Garza N.M., Chen Y., Watrous J.D., Murgia M., Trivedi P.P., Anderson S.K., Tomar D., Nilsson R., Madesh M., Jain M, & Gohil V.M. (2020). Yeast homologs of human MCUR1 regulate mitochondrial proline metabolism. Nature Communications, 11, 4866.

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Protein Expression Analysis using CN-PAGE and SDS-PAGE

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CN-PAGE was performed on conditioned media prepared on HEK293 cells expressing ^FTTTR variants using a 4%–16% gradient native PAGE gel (Life Technologies) as described previously (Chen et al., 2014 (link)). SDS-PAGE was performed on conditioned media or whole-cell lysates using 15% Tris-glycine gels. CN-PAGE and SDS-PAGE gels were transferred onto a nitrocellulose membrane (Bio-Rad) for immunoblotting. Membranes were blocked and then incubated with primary antibodies. Primary antibodies were then detected using appropriate IRDye-conjugated secondary antibodies and then analyzed using the Odyssey Infrared Imaging System (Li-COR Biosciences).

Chen J.J., Genereux J.C., Suh E.H., Vartabedian V.F., Rius B., Qu S., Dendle M.T., Kelly J.W, & Wiseman R.L. (2016). Endoplasmic Reticulum Proteostasis Influences the Oligomeric State of an Amyloidogenic Protein Secreted from Mammalian Cells. Cell chemical biology, 23(10), 1282-1293.

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