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- 2026年04月12日
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文献和实验Purification of Resistance Protein Complexes Using a Biotinylated Affinity (HPB) Tag
function. Many R proteins are associated with the plasma membrane (PM) and expressed at low levels. Here, we describe a method to purify such low-abundance PM R protein �complexes from Arabidopsis using a biotinylated affinity tag, called the HPB tag
Western Blotting with Biotinylated Antibodies
. 3.Centrifuge the homogenate (16,000 x g, 15 C) for 5 minutes to pellet insoluble material, then discard pellet. 4.Dilute an aliquot of the tissue lysate sample at least 10-fold for the BCA (Pierce) protein concentration assay. POLYACRYLAMIDE GEL
Selection of Antibodies Against Biotinylated Antigens
, but may also be problematic because the passive adsorption on plastic at pH 9.6 is a mechanism of protein denaturation. Under these conditions, 95% of adsorbed proteins are nonfunctional (2 ,3 ) This problem is not important for a classical enzyme-linked immunosorbant assay
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