![]() ![]() The sensitivity and rapidity of protein detection in QD-based approach is very high, with detection limits up to 10 pg of protein. The QD-conjugated antibodies are used to overcome the limitations of conventional western blot technique. The QDs are functionalized with antibodies of prostate apoptosis response-4 (Par-4), poly(ADP-ribose) polymerases and actin to specifically bind with the proteins localized in the nucleus and cytoplasm of the cells, respectively. High resolution transmission electron microscopy, Raman spectroscopy, fourier transform infrared spectroscopy, fluorescence spectroscopy and X-ray diffraction are used to characterize the properties of the quantum dots. Highly luminescent CdTe and (CdTe)ZnS QDs are synthesized by aqueous method. In the present study, we report a quantum dot (QD)-tailored western blot analysis for a sensitive, rapid and flexible detection of the nuclear and cytoplasmic proteins. Kale, Sonia [Agharkar Research Institute (India) Kale, Anup [University of Alabama, Center for Materials for Information Technology (United States) Gholap, Haribhau Rana, Abhimanyu [National Chemical Laboratory, Physical and Materials Chemistry Division (India) Desai, Rama [National Centre for Cell Science (India) Banpurkar, Arun [University of Pune, Department of Physics (India) Ogale, Satishchandra, E-mail: [National Chemical Laboratory, Physical and Materials Chemistry Division (India) Shastry, Padma, E-mail: [National Centre for Cell Science (India) Quantum dot bio-conjugate: as a western blot probe for highly sensitive detection of cellular proteinsĮnergy Technology Data Exchange (ETDEWEB) maxima that may present potential new drug targets or vaccine candidates for coccidiosis. Conclusions The data presented here suggest that specific genes identified between the two strains may be important molecules in the immunogenicity of E. The specific differentially expressed genes were finally verified by RT-PCR and qRT-PCR analyses. Further analysis found that six contigs from strain SH and three from strain NT shared significant identities with previously reported proteins, and one contig was presumed to be novel. Nucleotide sequencing analysis of these clones revealed ten specific contigs (six from strain SH and four from strain NT). Dot-blot hybridization revealed a total of 86 differentially expressed clones (63 from strain SH and 23 from strain NT). Results A total of 561 clones were selected from both cDNA libraries and the length of the inserted fragments was 0.25–1.0 kb. Two subtractive cDNA libraries were constructed using suppression subtractive hybridization (SSH) and specific genes were further analyzed by dot-blot hybridization and qRT-PCR analysis. maxima strains SH and NT, which were found to have significant differences in immunogenicity in our previous research. Methods Total RNA and mRNA were isolated from unsporulated oocysts of E. However, the genetic basis of these phenotypes remains unclear. Our method does not require any additional equipment or time compared to the conventional procedure with traditional fluo.Īnalysis of differentially expressed genes in two immunologically distinct strains of Eimeria maxima using suppression subtractive hybridization and dot-blot hybridizationīackground It is well known that different Eimeria maxima strains exhibit significant antigenic variation. We achieved a detection limit at the single-picogram level in dot blots with conventional Western blotting, we detected 50 pg of transferrin and trypsin inhibitor after SDS-PAGE and transfer onto a PVDF membrane. We demonstrate ultrasensitive fluorescence imaging of proteins on Western blots using a bright, compact, and orange-emitting semiconducting polymer dot (CN-PPV). Ultrasensitive Detection of Proteins on Western Blots with Semiconducting Polymer Dots ![]()
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