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The samples incubated with plain DMEM did not show any protein bands (data not shown).
Very weak luminescence bands (data not shown) were detected in the red-NIR region for both the yellows and ascribed to deep level emissions (DLE) [11]; these signals although too low to be used for confirming the stoichiometry, can be considered to further underline the similarity between the chemical composition of pastose and diluted Cd-based yellows.
When whole cell extracts from asynchronous HeLa cells were tested on Western blot, the antibody produced several cross-reacting bands (data not shown).
No available antibodies screened to date are able to detect endogenous PINK1 without having additional protein bands (data not shown), so qRT-PCR was used to confirm decreased PINK1 expression in the shRNA lines.
The antibody recognized the over-expressed proteins (∼72 kDa for EGFP-STK35) and a prominent band at about 58 kDa specifically whereas the pre-immune serum of the same rabbit did not detect these bands (data not shown).
Following the addition of 10 mM sodium dithionite to the incubation mixture a spectrum indicative of the ferrohaem-HmuY complex developed (a 422nm Soret and 526 and 556 nm Q bands) (data not shown).
Similar(25)
The anti-Lltryp2-peptide antibody revealed no band (data not shown).
Indicative of the presence of plasmids, we observed a faint band in CS-505 (Figure S2) plus a second band (data not shown) of approximately 30 kb.
Lambda phosphatase treatment of the RECQ1 immunoprecipitates in the presence of phosphatase inhibitors did not affect the mobility of the slower migrating band (data not shown).
In other experiments, SDS-PAGE electresolution resolution was improved by running a longer gel, revealing an additional 42 kDa band (data not shown).
Using both 5' and 3' fragments of OsPIE1 cDNA as the mixed probes in Northern blot, we detected a single, strongly hybridized band (data not shown) of 6.7 kb in size, consistent with its predicted length.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com