Fluoroquinolones are used to deal with severe bacterial infections, together with these brought on by Escherichia coli and Salmonella enterica. The emergence of plasmid-mediated quinolone resistance (PMQR) symbolize a brand new problem to the profitable therapy of Gram-negative infections. As a part of a long-term technique to generate a reference database of closed plasmids from antimicrobial resistant foodborne micro organism, we carried out long-read sequencing of 11 E. coli isolates from retail meats that had been non-susceptible to ciprofloxacin. Every of the isolates had PMQR genes, together with qnrA1, qnrS1, and qnrB19.
The 4 qnrB19 genes had been carried on two distinct ColE-type plasmids amongst isolates from pork chop and floor turkey and had been similar to plasmids beforehand recognized in Salmonella. Seven different plasmids differed from every other sequences in GenBank and comprised IncF and IncR plasmids that ranged in measurement from 48 to 180 kb. These plasmids additionally contained completely different mixtures of resistance genes, together with these conferring resistance to beta-lactams, macrolides, sulfonamides, tetracycline, and heavy metals.
Though comparatively few isolates have PMQR genes, the identification of various plasmids in a number of retail meat sources suggests the potential for additional unfold of fluoroquinolone resistance, together with by way of co-selection. These outcomes spotlight the worth of long-read sequencing in characterizing antimicrobial resistance genes of public well being concern. We additional present that these unusually excessive lactose uptake charges are poisonous to the cells, resulting in elevated cell leakiness and lysis. Lastly, we exhibit that in distinction to plasmid-based T7 expression methods, IPTG induction is useful for genome-integrated T7 expression methods regarding cell health and productiveness.
Identification of σE-Dependent Promoter Upstream of clpB from the Pathogenic Spirochaete Leptospira interrogans by Making use of an E. coli Two-Plasmid System.
There may be restricted info on gene expression within the pathogenic spirochaete Leptospirainterrogans and genetic mechanisms controlling its virulence. Transcription is step one in gene expression that’s typically decided by environmental results, together with infection-induced stresses. Alterations within the surroundings lead to important modifications within the transcription of many genes, permitting efficient adaptation of Leptospira to mammalian hosts. Thus, promoter and transcriptional begin web site identification are essential for figuring out gene expression regulation and for the understanding of genetic regulatory mechanisms current in Leptospira.
Right here, we characterised the promoter area of the L. interrogans clpB gene (clpBLi) encoding an AAA+ molecular chaperone ClpB important for the survival of this spirochaete beneath thermal and oxidative stresses, and likewise throughout an infection of the host. Primer extension evaluation demonstrated that transcription of clpB in L. interrogans initiates at a cytidine situated 41 bp upstream of the ATG initiation codon, and, to a lesser extent, at an adenine situated 2 bp downstream of the recognized web site. Transcription of each transcripts was heat-inducible.
Dedication of clpBLi transcription begin web site, mixed with promoter transcriptional exercise assays utilizing a modified two-plasmid system in E. coli, revealed that clpBLi transcription is managed by the ECF σE issue. Of the ten L. interrogans ECF σ components, the issue encoded by LIC_12757 (LA0876) is almost certainly to be the important thing regulator of clpB gene expression in Leptospira cells, particularly beneath thermal stress. Moreover, clpB expression could also be mediated by ppGpp in Leptospira.
Figuring out the Virulence Properties of Escherichia coli ST131 Containing Bacteriocin-Encoding Plasmids Utilizing Quick- and Lengthy-Learn Sequencing and Evaluating Them with These of Different E. coli Lineages.
Escherichia coli ST131 is a scientific problem as a consequence of its multidrug resistant profile and profitable international unfold. They’re typically related to difficult infections, significantly urinary tract infections (UTIs). Bacteriocins play an vital function to outcompete different microorganisms current within the human intestine. Right here, we characterised bacteriocin-encoding plasmids present in ST131 isolates of sufferers affected by a UTI utilizing each short- and long-read sequencing. Colicins Ia, Ib and E1, and microcin V, had been recognized amongst plasmids that additionally contained resistance and virulence genes.
To research if the potential transmission vary of the colicin E1 plasmid is influenced by the presence of a resistance gene, we constructed a pressure containing a plasmid which had each the colicin E1 and blaCMY-2 genes. No distinction in transmission vary was discovered between transformant and wild-type strains. Nonetheless, a statistically considerably distinction was present in adhesion and invasion capacity. Bacteriocin-producing isolates from each ST131 and non-ST131 lineages had been capable of inhibit the expansion of different E. coli isolates, together with different ST131. In abstract, plasmids harboring bacteriocins give extra benefits for extremely virulent and resistant ST131 isolates, enhancing the power of those isolates to compete with different microbiota for a distinct segment and thereby rising the danger of an infection.
The current examine aimed to judge the affect of induction situations (IPTG focus, temperature, and induction time) on the plasmid pQE-30 stability and 503 antigen expression of Leishmania i. chagasi in Escherichia coli M15. Batch cultures had been carried out at 37 °C and induced by the addition of various IPTG concentrations (0.01 to 1.5 mM). Subsequently, experiments had been carried out at completely different temperatures (27 to 42 °C), evaluating the affect of induction time (0.5 to six h after the beginning of the tradition). The outcomes confirmed that IPTG toxicity brought on a metabolic stress within the cells and, consequently, the microorganism development decreased.
PRDM13 Polyclonal Antibody |
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A-2013 | EpiGentek |
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PRDM16 Polyclonal Antibody |
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A-2016 | EpiGentek |
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PRDM17 Polyclonal Antibody |
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A-2017 | EpiGentek |
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EZH1 Polyclonal Antibody |
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A-2018 | EpiGentek |
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HSF1 Polyclonal Antibody |
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A-2401 | EpiGentek |
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PRMT3 Polyclonal Antibody |
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A-3003 | EpiGentek |
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SET1 Polyclonal Antibody |
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A-3011 | EpiGentek |
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SET07 Polyclonal Antibody |
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A-3013 | EpiGentek |
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LSD1 Polyclonal Antibody |
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A-3018 | EpiGentek |
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HDAC10 Polyclonal Antibody |
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A-4010 | EpiGentek |
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PCAF Polyclonal Antibody |
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A-4012 | EpiGentek |
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Histone H3K9ac (Acetyl H3K9) Polyclonal Antibody |
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A-4022 | EpiGentek |
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Histone H3K14ac (Acetyl H3K14) Polyclonal Antibody |
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A-4023 | EpiGentek |
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Histone H3K18ac (Acetyl H3K18) Polyclonal Antibody |
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A-4024 | EpiGentek |
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Histone H3K23ac (Acetyl H3K23) Polyclonal Antibody |
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A-4025 | EpiGentek |
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Histone H3K56ac (Acetyl H3K56) Polyclonal Antibody |
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A-4026 | EpiGentek |
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Histone H3K4me1 (H3K4 Monomethyl) Polyclonal Antibody |
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A-4031 | EpiGentek |
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Histone H3K4me2 (H3K4 Dimethyl) Polyclonal Antibody |
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A-4032 | EpiGentek |
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Histone H3K4me3 (H3K4 Trimethyl) Polyclonal Antibody |
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A-4033 | EpiGentek |
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Histone H3K9me1 (H3K9 Monomethyl) Polyclonal Antibody |
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A-4034 | EpiGentek |
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Histone H3K9me2 (H3K9 Dimethyl) Polyclonal Antibody |
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A-4035 | EpiGentek |
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Histone H3K9me3 (H3K9 Trimethyl) Polyclonal Antibody |
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A-4036 | EpiGentek |
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Histone H3K27me1 (H3K27 Monomethyl) Polyclonal Antibody |
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A-4037 | EpiGentek |
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Histone H3K27me2 (H3K27 Dimethyl) Polyclonal Antibody |
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A-4038 | EpiGentek |
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Histone H3K27me3 (H3K27 Trimethyl) Polyclonal Antibody |
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A-4039 | EpiGentek |
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Histone H3K36me1 (H3K36 Monomethyl) Polyclonal Antibody |
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A-4040 | EpiGentek |
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Histone H3K36me2 (H3K36 Dimethyl) Polyclonal Antibody |
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A-4041 | EpiGentek |
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Histone H3K36me3 (H3K36 Trimethyl) Polyclonal Antibody |
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A-4042 | EpiGentek |
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Histone H3K79me1 (H3K79 Monomethyl) Polyclonal Antibody |
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A-4043 | EpiGentek |
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Histone H3K79me2 (H3K79 Dimethyl) Polyclonal Antibody |
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A-4044 | EpiGentek |
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Histone H3K79me3 (H3K79 Trimethyl) Polyclonal Antibody |
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A-4045 | EpiGentek |
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Histone H4K20me1 (H4K20 Monomethyl) Polyclonal Antibody |
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A-4046 | EpiGentek |
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Histone H4K20me2 (H4K20 Dimethyl) Polyclonal Antibody |
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A-4047 | EpiGentek |
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Histone H4K20me3 (H4K20 Trimethyl) Polyclonal Antibody |
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A-4048 | EpiGentek |
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Histone H3K27ac (Acetyl H3K27) Polyclonal Antibody |
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A-4708 | EpiGentek |
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CRISPR Cas9 Monoclonal Antibody [7A9] |
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A-9000 | EpiGentek |
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CRISPR/Cas9 (SaCas9) Monoclonal Antibody [6H4] |
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A-9001 | EpiGentek |
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CLIMP-63 Polyclonal Antibody |
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A-0701 | EpiGentek |
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DNMT3A Polyclonal Antibody |
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A-1003 | EpiGentek |
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DNMT3B Polyclonal Antibody |
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A-1004 | EpiGentek |
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DNMT3L Polyclonal Antibody |
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A-1005 | EpiGentek |
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MBD3 Polyclonal Antibody |
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A-1008 | EpiGentek |
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MGMT Polyclonal Antibody |
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A-1010 | EpiGentek |
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MeCP2 Polyclonal Antibody |
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A-1012 | EpiGentek |
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TET1 Polyclonal Antibody |
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A-1020 | EpiGentek |
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DNMT1 Polyclonal Antibody |
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A-1700 | EpiGentek |
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TET2 Polyclonal Antibody |
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A-1701 | EpiGentek |
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MBD2 Polyclonal Antibody |
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A-1713 | EpiGentek |
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PRDM2 Polyclonal Antibody |
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A-2002 | EpiGentek |
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PRDM3 Polyclonal Antibody |
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A-2003 | EpiGentek |
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PRDM6 Polyclonal Antibody |
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A-2006 | EpiGentek |
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PRDM12 Polyclonal Antibody |
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A-2012 | EpiGentek |
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PRDM14 Polyclonal Antibody |
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A-2014 | EpiGentek |
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EZH2 Polyclonal Antibody |
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A-2019 | EpiGentek |
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EED Polyclonal Antibody |
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A-2020 | EpiGentek |
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Swi2/SNF2 Polyclonal Antibody |
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A-2023 | EpiGentek |
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SNFa/BRM Polyclonal Antibody |
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A-2025 | EpiGentek |
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Ini1 Polyclonal Antibody |
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A-2026 | EpiGentek |
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RNA Polymerase II Monoclonal Antibody [CTD4H8] |
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A-2032 | EpiGentek |
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CBX5 Polyclonal Antibody |
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A-2701 | EpiGentek |
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EGLN1 Polyclonal Antibody |
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A-2702 | EpiGentek |
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RBBP4 Polyclonal Antibody |
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A-2703 | EpiGentek |
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SIN3A Polyclonal Antibody |
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A-2704 | EpiGentek |
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CTBP1 Polyclonal Antibody |
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A-2705 | EpiGentek |
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PADI4 Polyclonal Antibody |
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A-2706 | EpiGentek |
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SMARCA5 Polyclonal Antibody |
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A-2707 | EpiGentek |
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IDH1 Polyclonal Antibody |
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A-2708 | EpiGentek |
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CHD4 Polyclonal Antibody |
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A-2709 | EpiGentek |
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CHAF1A Polyclonal Antibody |
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A-2710 | EpiGentek |
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BRD7 Polyclonal Antibody |
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A-2712 | EpiGentek |
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UHRF1 Polyclonal Antibody |
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A-2714 | EpiGentek |
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UHRF2 Polyclonal Antibody |
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A-2715 | EpiGentek |
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SMARCE1 Polyclonal Antibody |
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A-2716 | EpiGentek |
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ACTL6A Polyclonal Antibody |
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A-2717 | EpiGentek |
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MPG Polyclonal Antibody |
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A-2718 | EpiGentek |
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HIF1AN Polyclonal Antibody |
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A-2719 | EpiGentek |
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RNF2 Polyclonal Antibody |
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A-2720 | EpiGentek |
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PCGF6 Polyclonal Antibody |
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A-2721 | EpiGentek |
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SMARCB1 Polyclonal Antibody |
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A-2722 | EpiGentek |
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ALKBH3 Polyclonal Antibody |
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A-2723 | EpiGentek |
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BBOX1 Polyclonal Antibody |
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A-2724 | EpiGentek |
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BTAF1 Polyclonal Antibody |
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A-2725 | EpiGentek |
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CXXC1 Polyclonal Antibody |
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A-2726 | EpiGentek |
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KDM4B Polyclonal Antibody |
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A-2727 | EpiGentek |
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HELLS Polyclonal Antibody |
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A-2728 | EpiGentek |
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ING3 Polyclonal Antibody |
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A-2729 | EpiGentek |
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ING4 Polyclonal Antibody |
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A-2730 | EpiGentek |
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PHC1 Polyclonal Antibody |
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A-2731 | EpiGentek |
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SMARCAD1 Polyclonal Antibody |
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A-2732 | EpiGentek |
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USP16 Polyclonal Antibody |
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A-2733 | EpiGentek |
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SMARCC1 Polyclonal Antibody |
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A-2734 | EpiGentek |
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ERCC6L Polyclonal Antibody |
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A-2735 | EpiGentek |
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SMYD5 Polyclonal Antibody |
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A-2736 | EpiGentek |
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ATF2 Polyclonal Antibody |
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A-2738 | EpiGentek |
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BMI1 Polyclonal Antibody |
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A-2739 | EpiGentek |
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CTBP2 Polyclonal Antibody |
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A-2740 | EpiGentek |
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CTCF Polyclonal Antibody |
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A-2741 | EpiGentek |
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MTA2 Polyclonal Antibody |
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A-2742 | EpiGentek |
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PMS1 Polyclonal Antibody |
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A-2744 | EpiGentek |
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SFN Polyclonal Antibody |
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A-2745 | EpiGentek |
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SMARCA4 Polyclonal Antibody |
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A-2746 | EpiGentek |
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SMARCAL1 Polyclonal Antibody |
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A-2747 | EpiGentek |
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SMARCC2 Polyclonal Antibody |
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A-2748 | EpiGentek |
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MuERVL-Gag Polyclonal Antibody |
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A-2801 | EpiGentek |
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PRMT1 Polyclonal Antibody |
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A-3001 | EpiGentek |
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PRMT2 Polyclonal Antibody |
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A-3002 | EpiGentek |
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PRMT4 Polyclonal Antibody |
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A-3004 | EpiGentek |
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PRMT5 Polyclonal Antibody |
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A-3005 | EpiGentek |
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PRMT7 Polyclonal Antibody |
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A-3007 | EpiGentek |
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SUV39H2 Polyclonal Antibody |
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A-3010 | EpiGentek |
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SET9 Polyclonal Antibody |
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A-3014 | EpiGentek |
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The induction with IPTG may additionally be related to the plasmid pQE-30 instability, as a consequence of metabolic burden imposed by the recombinant protein expression. The optimum situations for 503 antigen expression of Leishmania i. chagasi in Escherichia coli M15 had been an IPTG focus of 1.Zero mM, temperature of 37 °C, and induction time of two h. The utmost antigen focus obtained was 0.119 ± 0.009 g/L, about seven occasions greater than the bottom focus. Due to this fact, the outcomes confirmed that 503 antigen will be produced in laboratory; nonetheless, it requires extra research to attenuate the plasmid instability and enhance to industrial scale.