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. 2007 May 1;364(1):92-4.
doi: 10.1016/j.ab.2007.01.019. Epub 2007 Jan 19.

Fluorescence-based analysis of aminoacyl- and peptidyl-tRNA by low-pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Robert N Kirchdoerfer  1 , Joseph J-T Huang, Molly K Isola, Silvia Cavagnero
Affiliations

Fluorescence-based analysis of aminoacyl- and peptidyl-tRNA by low-pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Robert N Kirchdoerfer et al. Anal Biochem. .
No abstract available

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Figures

Fig. 1
Fig. 1
Analysis of BODIPYFL-Met-tRNA fMet incubated under different conditions by low pH TA SDS-PAGE. BODIPYFL-Met-tRNAfMet samples were hydrolyzed in 0.1 M tris buffers, adjusted to pH values ranging from 6. to 9 by addition of aqueous HCl, at 37 °C for 1.5 hrs. Bands corresponding to BODIPYFL-Met-tRNAfMet and its ester bond hydrolysis products are enclosed in green and orange rectangles, respectively.
Fig. 2
Fig. 2
(A) Analysis of cell-free-generated peptidyl-tRNAs bearing N-terminal portions of the amino acid sequence of apoMb by low pH TA (Lanes 1, 2, 5 and 6) and reference pH TT (Lanes 3, 4, 7, and 8) SDS PAGE. All ribosome-associated nascent apoMb polypeptides were N-terminally labeled with BODIPYFL-Met-tRNAfMet. Bands corresponding to nascent polypeptides covalently linked to tRNA and tRNA-released by treatment with puromycin are enclosed in green and orange rectangles, respectively. (B) Diagram providing the quantitative extent of ester hydrolysis for peptidyl-tRNA run on the TA and TT gels in panel A (lanes 1, 3, 5, and 7). For each lane, the percent of ester hydrolysis was determined as the ratio of hydrolyzed product to total fluorescence intensity.
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