|
| Status |
Public on Feb 21, 2022 |
| Title |
aptamer m12-3773 replicate 2 |
| Sample type |
protein |
| |
|
| Source name |
aptamer library selected on human islet
|
| Organism |
Homo sapiens |
| Characteristics |
target: unknonw protein expressed on human islets
|
| Extracted molecule |
protein |
| Extraction protocol |
NA
|
| Label |
Alexafluor647
|
| Label protocol |
5'Biotinylated aptamers were conjugated overnight at 4C with Alexafluor 647 labelled straptavidin (Biolegend) 4:1 molar ratio. The conjugates were purified with an Amicon 50,000 MWCO - (Millipore) spin filter.
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| |
|
| Hybridization protocol |
HuProt 2.0 arrays (ArrayIt) were prepared for binding as recommended by the manufacturer. The protein surface of the array was deactivated with 3 ml of “Chem block “buffer (Arrayit) for one hour at room temperature. After deactivation, the array was blocked with blocking buffer (Blockit - Arrayit, supplemented to 450µg/ml yeast RNA - Ambion) for one hour and then hybridized for 30’ at 4ºC with the streptavidin aptamer-conjugate (20 picomoles) in 200µl of blocking buffer. Then, the arrays were washed (5 x 5’) in washing buffer (0.01 % Tween 20 - Biorad in 0.5mM Mg2+ PBS), and dried using a Microarray centrifuge (ArrayIt).
|
| Scan protocol |
Array have been analyzed on a genepix 4000B microarray reader.
|
| Description |
Islet14 replicate B
|
| Data processing |
Protein arrays have been analyzed using the PAA package (v. 1.0)(Turewicz et al, 2016) in R (v. 3.4.4); before preprocessing, each microarray has been visually inspected for any artifact using the plotArray function. Then, the raw intensity signals were extracted from the .gpr files and B11the median fluorescence intensities at 635nm wavelength was imputed as raw intensity foreground signals; specifically, intensity signals of replicated spots were mean centered and background corrected using the default parameters, e.g. the saddle variant of the normexpr method in the backgroundCorrect function, and were finally normalized using the quantile method of the normalizeArrays function.
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| |
|
| Submission date |
Nov 27, 2020 |
| Last update date |
Feb 21, 2022 |
| Contact name |
Silvio Bicciato |
| E-mail(s) |
silvio.bicciato@unipd.it
|
| Phone |
+39-049-827-6108
|
| Organization name |
University of Padova
|
| Department |
Molecular Medicine
|
| Street address |
via U. Bassi 59/b
|
| City |
Padova |
| ZIP/Postal code |
35131 |
| Country |
Italy |
| |
|
| Platform ID |
GPL29450 |
| Series (1) |
| GSE162273 |
RNA aptamers specific for transmembrane p24 trafficking protein 6 and Clusterin for the targeted delivery of imaging reagents and RNA therapeutic to human β cells |
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