Infected cells were labeled with 0.1 mCi (3.7 MBq) per well of [35S]Cys-Met mix (Hartmann Analytic). After the respective labeling time, the cells were lysed, divided in two samples, and mixed with protein A-agarose beads (Santa Cruz) and the respective antibody. After overnight incubation at 4°C, the proteins were eluted and separated on Mini-Protean 10% TGX gels (Bio-Rad). Dried gels were exposed to Kodak cassette-K BaFBr:Eu phosphor screens overnight and imaged using the Bio-Rad Personal Molecular Imager system. Bands were quantified using the Bio-Rad Quantity One 1-D Analysis software package.
Quantity one 1 d analysis software package
Manufactured by Bio-Rad
Sourced in United States
Quantity One 1-D analysis software package is a comprehensive software solution designed for the analysis and quantification of 1-dimensional electrophoresis gels and blots. The software provides tools for image capture, analysis, and data management.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using quantity one 1 d analysis software package
1
Immunoprecipitation and Autoradiography
2
Leptospira DNA Extraction and Amplification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The genomic DNA of Leptospira was extracted by phenol-chloroform method (Merck, Germany). The final volume (25 µL) of each reaction mixture contained 2.5 µL of 1X PCR buffer (SinaClone, Iran), 0.5 mM of 10 mM deoxynucleoside triphosphate (SinaClone, Iran), 1 µM of 10 pmol each corresponding primer, 0.75 of 50 mM MgCl2 (SinaClone, Iran), 0.3 unit/µL of 0.5 unit Taq DNA polymerase (SinaClone, Iran) and 1 µL of 100 macro gram DNA template. Amplification was achieved under the following conditions: 1 denaturation cycle at 93˚C for 5 minutes; 35 cycles of denaturation at 93˚C for 1 minute, annealing at 54˚C for 1 minute, elongation at 72˚C for 1 minute, and a final elongation at 72˚C for 10 minutes (14 (link)). The amplified products were analyzed by 2% agarose gel electrophoresis and allelic sizes estimated by Quantity One 1-D analysis software package (Bio-Rad, USA). The size of the amplified products was estimated by comparison with a 100 bp plus ladder (15 (link), 16 (link)). Some of the amplified products were sequenced by Macrogene Company in South Korea.
3
Western Blot Analysis of Colon ILC2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Colon ILC2 cells were lysed with cold RIPA buffer (Thermo Fisher Scientific, Waltham, MA, USA) for protein extraction. Totally 25 µg protein was separated by 12% SDS-PAGE, which was then transferred onto the polyvinylidene fluoride membrane. The membrane was blocked with 5% nonfat milk at room temperature for 2 h, and further incubated overnight with rabbit anti-Arginase1 (anti-Arg1) (Sangon Biotech, Shanghai, China), rabbit anti-Killer cell lectin-like receptor G1 (KLRG1) (Abcam, Cambridge, MA, USA), rabbit anti-receptor B (IL-17RB) (Solarbio, Beijing, China), or mouse anti-β-actin antibodies (Abcam, Cambridge, MA, USA) at 4 °C. After washing, the membranes were incubated with the secondary antibody (Abcam, Cambridge, MA, USA) and finally processed using the enhanced chemiluminescence reaction kit (Cell Signaling Technology, Danvers, MA, USA). The relative expression levels were quantified using the Quantity One 1-D analysis software package (Bio-Rad, Hercules, CA, USA).
4
Microbial Diversity in Dental Implants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Quantity One 1D-Analysis Software package (v4.6.5, Bio-Rad) was used for the evaluation of the individual 16S rDNA banding patterns.
The statistical analysis compared the microbial diversity of sulcus fluid around implants to the remaining dentition and the null hypothesis is rejected if a significant difference is detected between implants and remaining dentition.
The null hypothesis is:
Data documentation and evaluation was performed with the data processing program SPSS/PC Version 20.0 for Windows (SPSS, Chicago, IL, USA).
The band migration patterns within individual patients were also compared. Bands occurring at the same height (±4%) within the gels were assigned as belonging to the same bacterial species (low diversity), whereas differences in the observed patterns are indicative of an altered microbial community composition (high diversity).
The statistical analysis compared the microbial diversity of sulcus fluid around implants to the remaining dentition and the null hypothesis is rejected if a significant difference is detected between implants and remaining dentition.
The null hypothesis is:
-H0 (1): No difference in microbial diversity between implants and the remaining dentition.
-HA(1): Significant difference in microbial diversity between implants and the remaining dentition.
Data documentation and evaluation was performed with the data processing program SPSS/PC Version 20.0 for Windows (SPSS, Chicago, IL, USA).
The band migration patterns within individual patients were also compared. Bands occurring at the same height (±4%) within the gels were assigned as belonging to the same bacterial species (low diversity), whereas differences in the observed patterns are indicative of an altered microbial community composition (high diversity).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!