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These were found to be Val-111 (top band, cleavage site LC↓VC), Gln-131 (middle band, cleavage site LC↓QL), and Leu-150 (lower band, cleavage site IV↓LQ), resulting in theoretical molecular masses of 41.5, 39.5, and 37.2 kDa, respectively.
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In contrast to the presence of several discrete bands in the final purified sample of isolated wild-type hFasRECD part [ 15], this mutant gave a single band after cleavage with bovine thrombin due to the lack of heterologous N-glycosylation.
The defined size of the star bands from cleavage reactions of λ DNA suggests that the DraIII star activity is not random and has certain specificity.
The difference in size between the two bands suggested cleavage between amino acids 225 and 230.
Activities of PARP and caspase-3 were measured as the proportion of cleavage band intensity to the total bands and were calculated as follows: % PARP or caspase-3 = 100% × Tc/Tt, where Tc is the intensity value of the cleavage bands and Tt is the intensity value of total bands.
With their shoulder straps thin as ribbon and narrow back bands, the cleavage-baring bras resemble two clam-shell halves looped together with string (similar to what the heroine wears in "The Little Mermaid").
Further Western blot experiments using a protocol optimized to improve size discrimination across this mass range yielded a band for the cleavage product at ∼30.1 kDa (Figure S9b).
The number of rectangles or circles corresponds to the intensity of the cleavage band at that position, with more symbols indicating a higher probability of "single-strandedness".
As can be seen in Figure 2a we detected a faint cleavage band for both caspases as soon as 5 h of culture in suspension.
We also observed a band consistent with cleavage of GFP from the GFP LGG-1 fusion protein that was similarly sensitive to bec-1(RNAi).
The dissolved egg envelope yielded a 37 kDa band caused by cleavage of the N-ZPd site in ZPC and two low molecular bands at 17 and 16 kDa produced by cleavage mid-ZPd in ZPB [ 7].
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