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tRNA Probes

Photo-crosslinking aa–tRNAs

 Upon illumination with 320-370 nm light, the photo-reactive probes below  form highly reactive species: ANB (5-azido-2-nitrobenzoyl-) forms a nitrene, AP (4,4-azipentanoyl-) forms a carbene, and BP (benzophenone-) forms a triplet biradical.  After incorporation into a protein during in vitro translation, a probe will photo-crosslink to whatever molecular species is in close proximity.  The resulting photo-adduct is analyzed to identify the protein's binding or assembly partners in a particular sample.

Photo-crosslinking targets can be easily purified using a BAD photo-reactive and affinity transfer probe that contains biotin, arylazide, and disulfide moieties.  After a BAD-labeled protein is photo-crosslinked to the target protein or nucleic acid, reduction of the disulfide bond separates the target from the BAD-labeled protein, but the biotin remains covalently attached to the target.  The biotin-containing target can then be separated from the rest of the sample using avidin- or streptavidin-containing materials to facilitate subsequent analysis and identification.

Each tube contains 400 pmoles of aa-tRNA that have been dialyzed into 25-30 μl of 1 mM KOAc (pH 5.0), 2 mM Mg(OAc)2.

Item #

Photo-crosslinking  aa–tRNAs

Quantity per tube

Cost per tube

A-12

εANB-[14C]Lys-tRNAamb

400 pmoles

$ 75
A-13

 εBP-[14C]Lys-tRNAamb

400 pmoles

$ 75
A-14 εAP-[14C]Lys-tRNAamb 400 pmoles $ 80
A-52

non-RA  εANB-Lys-tRNAamb

400 pmoles $ 75
A-53

non-RA  εBP-Lys-tRNAamb

400 pmoles $ 75
 A-54

non-RA  εAP-Lys-tRNAamb

400 pmoles $ 75
       
A-504 non-RA  εBAD-Lys-tRNAamb 400 pmoles $ 75
 
C-13

E. coli  BP-S-[14C]Cys-tRNACys

400 pmoles

$ 85
C-53

non-RA  E. coli  BP-S-Cys-tRNACys

400 pmoles $ 80
 
       
C-63 non-RA  yeast  BP-S-Cys-tRNACys 400 pmoles $ 80
 
L-12

E. coli  εANB-[14C]Lys-tRNALys

400 pmoles

$ 70
L-13

E. coli  εBP-[14C]Lys-tRNALys

400 pmoles

$ 70
L-14

E. coli  εAP-[14C]Lys-tRNALys

400 pmoles
$ 70
L-52

non-RA E. coli  εANB-Lys-tRNALys

400 pmoles $ 70
L-53

non-RA  E. coli  εBP-Lys-tRNALys

400 pmoles $ 70
       
 
L-32

yeast  εANB-[14C]Lys-tRNALys

400 pmoles

$ 80
L-33

yeast  εBP-[14C]Lys-tRNALys

400 pmoles

$ 80
L-34

yeast  εAP-[14C]Lys-tRNALys

400 pmoles $ 80
L-62 non-RA  yeast  εANB-Lys-tRNALys 400 pmoles $ 80
L-63 non-RA  yeast  εBP-Lys-tRNALys 400 pmoles $ 80