HBV capsids were mixed with 5X SDS-loading dye (5% β-Mercaptoethanol, 0.02% Bromophenol blue, 30% Glycerol, 10% SDS, 250 mM Tris-HCl, PH 6.8) and boiled for 10 min before loading onto polyacrylamide-bound Mn2+-Phos-tag SDS-PAGE (Wako Pure Chemical Industries, Richmond, VA, USA)62 (link)63 64 (link). After electrophoresis, the gel was soaked for 10 min in protein transfer buffer (25 mM Tris, 192 mM glycine, 10% methanol) containing 1 mM EDTA, followed by soaking for another 10 min in the same buffer without EDTA. Western blot analysis was performed by using rabbit anti-HBc polyclonal antibody (DAKO, Glostrup, Denmark).
Mn2 phos tag sds page
Manufactured by Fujifilm
Sourced in United States
Mn2+-Phos-tag SDS-PAGE is a laboratory equipment product that enables the separation and detection of phosphorylated proteins in a sample. It utilizes a unique Phos-tag compound that binds to phosphate groups, allowing for the separation of phosphorylated and non-phosphorylated proteins during electrophoresis.
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Lab products found in correlation
2 protocols using mn2 phos tag sds page
1
HBV Capsid Analysis via Phos-tag SDS-PAGE
2
In Vitro Phosphorylation of Lgl by aPKC
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A reaction mixture containing 15 μM MBP-Lgl
647–673, the aPKC kinase domain, and 1 mM ATP in the reaction
buffer described above was set up and incubated at 30 °C for
90 min. Small aliquots of the reaction mixture were quenched via sodium
dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE)
loading buffer periodically during the incubation. The quenched samples
were run on Mn2+-Phos-tag SDS–PAGE (Wako USA). The
phosphorylated Lgl was detected on a Western blot using mouse anti-MBP
(1:1000, Santa Cruz Biotechnology) and bovine anti-mouse HRP (1:2000,
Santa Cruz Biotechnology) antibodies.
647–673, the aPKC kinase domain, and 1 mM ATP in the reaction
buffer described above was set up and incubated at 30 °C for
90 min. Small aliquots of the reaction mixture were quenched via sodium
dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE)
loading buffer periodically during the incubation. The quenched samples
were run on Mn2+-Phos-tag SDS–PAGE (Wako USA). The
phosphorylated Lgl was detected on a Western blot using mouse anti-MBP
(1:1000, Santa Cruz Biotechnology) and bovine anti-mouse HRP (1:2000,
Santa Cruz Biotechnology) antibodies.
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