State-of-the-art housekeeping proteins for quantitative western blotting: Revisiting the first draft of the human proteome

PROTEOMICS ◽  
2016 ◽  
Vol 16 (13) ◽  
pp. 1863-1867 ◽  
Author(s):  
Hyun-Gwan Lee ◽  
Jihoon Jo ◽  
Hyun-Hee Hong ◽  
Kee K. Kim ◽  
Joong-Ki Park ◽  
...  
Keyword(s):  
Western Blotting ◽  
State Of The Art ◽  
Human Proteome ◽  
Quantitative Western Blotting

scholarly journals Replacement of Vegetative ςA by Sporulation-Specific ςF as a Component of the RNA Polymerase Holoenzyme in Sporulating Bacillus subtilis

1999 ◽  
Vol 181 (8) ◽  
pp. 2346-2350 ◽  
Author(s):  
Matthew Lord ◽  
Daniela Barillà ◽  
Michael D. Yudkin
Keyword(s):  
Bacillus Subtilis ◽  
Rna Polymerase ◽  
Western Blotting ◽  
Sigma Factor ◽  
Sigma Factors ◽  
Relative Affinity ◽  
Rna Polymerase Holoenzyme ◽  
Quantitative Western Blotting

ABSTRACT Soon after asymmetric septation in sporulating Bacillus subtilis cells, ςF is liberated in the prespore from inhibition by SpoIIAB. To initiate transcription from its cognate promoters, ςF must compete with ςA, the housekeeping sigma factor in the predivisional cell, for binding to core RNA polymerase (E). To estimate the relative affinity of E for ςA and ςF, we made separate mixtures of E with each of the two sigma factors, allowed reconstitution of the holoenzyme, and measured the concentration of free E remaining in each mixture. The affinity of E for ςF was found to be about 25-fold lower than that for ςA. We used quantitative Western blotting to estimate the concentrations of E, ςA, and ςF in sporulating cells. The cellular concentrations of E and ςA were both about 7.5 μM, and neither changed significantly during the first 3 h of sporulation. The concentration of ςF was extremely low at the beginning of sporulation, but it rose rapidly to a peak after about 2 h. At its peak, the concentration of ςF was some twofold higher than that of ςA. This difference in concentration cannot adequately account for the replacement of ςA holoenzyme by ςF holoenzyme in the prespore, and it seems that some further mechanism—perhaps the synthesis or activation of an anti-ςA factor—must be responsible for this replacement.

Determination of Copy Number of c-Myc Protein per Cell by Quantitative Western Blotting

1999 ◽  
Vol 269 (1) ◽  
pp. 66-71 ◽  
Author(s):  
Claudia Rudolph ◽  
Gerold Adam ◽  
Andreas Simm
Keyword(s):  
Western Blotting ◽  
Copy Number ◽  
Quantitative Western Blotting

scholarly journals Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting

10.3791/59438 ◽  
2019 ◽  
Author(s):  
Yu-Ting Huang ◽  
Dinja van der Hoorn ◽  
Leire M. Ledahawsky ◽  
Anna A. L. Motyl ◽  
Crispin Y. Jordan ◽  
...  
Keyword(s):  
Western Blotting ◽  
Protein Levels ◽  
Quantitative Western Blotting
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