|
| Status |
Public on Jun 25, 2019 |
| Title |
HC 114 |
| Sample type |
protein |
| |
|
| Source name |
Culture supernatant from Mtb-stimulated whole blood
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Whole blood
histology: Healthy
|
| Treatment protocol |
Whole blood (250 µl) collected from participants using an EDTA anticoagulation tube was cultured in serum-free medium (DKW34-EHK0100, DAKEWE) in absence or presence of 2.5 µg/ml phytohemagglutinin (PHA, L8754, Sigma) or 10 µg/ml Mtb-antigen peptide pool for 24 h at 37°C, 5% CO2, and then the supernatant was collected.
|
| Extracted molecule |
protein |
| Extraction protocol |
Culture supernatant from Mtb-stimulated whole blood
|
| Label |
AlexaFluor 555
|
| Label protocol |
AlexaFluor 555-conjugated streptavidin was then added and incubated for 1 hour at room temperature.
|
| |
|
| Hybridization protocol |
2-fold diluted culture supernatant was added to each well. After overnight incubation at 4°C and extensive washing, the biotin-labeled detection antibody was added for 2 hours and then washed away. AlexaFluor 555-conjugated streptavidin was then added and incubated for 1 hour at room temperature.
|
| Scan protocol |
The signals (532 nm excitation, 635 nm emission) were scanned and extracted using an InnoScan 300 scanner (Innopsys, Carbonne, France).
|
| Description |
A combination of 16 non-overlapping arrays to quantitatively measure 640 human cytokines. Each antibody, together with the positive controls is arrayed in quadruplicate.
|
| Data processing |
Individual array spots were background subtracted locally by subtracting the median background across spot replicated in each sample, and then normalized using two positive controls. For protein quantification, standard curves were generated using purified proteins representing the proteins-of-interest.
|
| |
|
| Submission date |
Jun 24, 2019 |
| Last update date |
Jun 25, 2019 |
| Contact name |
Huihua Zhang |
| Organization name |
Jinan University
|
| Department |
Biomedicine Institute & Cell Biology Department
|
| Street address |
601, west Huangpu avenue, Tianhe district
|
| City |
Guangzhou |
| ZIP/Postal code |
510632 |
| Country |
China |
| |
|
| Platform ID |
GPL26838 |
| Series (1) |
| GSE133249 |
Identification of an eight-protein biosignature for the diagnosis of tuberculosis |
|
