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The procedures for the expression of mouse BCL-XL was described previously.
The procedures for the expression of proteins in Escherichia coli and subsequent purification via nickel column chromatography were described previously (Uppalapati et al., 2009; Shastry and Hancock, 2010).
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The objective of this study was to develop a reliable procedure for the expression of a full-length DGAT1 in E. coli using tung tree DGAT1 (one of the two isoforms from tung tree) as the model protein.
Cells were stained with combinations of fluorochrome-labelled Abs according to standard procedures for measuring the expression of surface or intracellular markers on FACS-Canto or FACS-Calibur flow cytometers (BD Biosciences).
RNAseIII-deficient Escherichia coli strain HT115 (rnc14:: ΔTn10) [32] was grown in LB broth for genetic engineering procedures or 2YT broth for the expression of dsRNA, both containing antibiotics ampicillin (100 µg/ml) and tetracycline (10 µg/ml).
With further development of antibodies and refinement of staining procedure, we carried out immunohistochemical analysis for the expression of GATA4 and GATA6 in a panel of ovarian non-neoplastic and cancer tissues in tissue microarray (Table 1).
For cDNA arrays, normalized expression ratios of 9050 genes were calculated, and the same procedure was applied to long-oligonucleotide arrays for the expression of 20,799 genes.
This test procedure resulted in a highly significant difference (p < 0.01) for the expression of BAX and a borderline significance (p = 0.06866) for the expression of p21.
Testing for the expression of CPP or CPA was conducted on day 10 using the pretest procedure.
The procedure for the expression and purification of recombinant 6xHis-SUMO fusion proteins was performed as follows.
We have established a uniform procedure for the expression and purification of the cyclin-dependent kinases CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1.
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