|
| Status |
Public on Aug 15, 2024 |
| Title |
Patient_075_After [PAT_075_A_IgA] |
| Sample type |
protein |
| |
|
| Source name |
Gonococcal infection
|
| Organism |
Homo sapiens |
| Characteristics |
Sex: Male
subject group: Individual with confirmed infection
tissue: Blood
tissue subtype: Serum
|
| Extracted molecule |
protein |
| Extraction protocol |
Human sera collected from the KEMRI clinics in Mtwapa and Malindi and from clinical trial NCT 04297436
|
| Label |
Goat Anti-Human IgA alpha chain (DyLight® 650) pre-adsorbed secondary antibody
|
| Label protocol |
Secondary antibodies were purchased from Abcam: Goat Anti-Human IgG Fc (DyLight® 650) pre-adsorbed secondary antibody (ab98622, Abcam, UK) and Goat Anti-Human IgA alpha chain (DyLight® 650) pre-adsorbed secondary antibody (ab96998, Abcam, UK)
|
| |
|
| Hybridization protocol |
Slides were blocked with 200 µL of SuperG Blocking Buffer (Grace Biolabs, US) per well and incubated for one hour at room temperature (RT). Washes were done three times with 300μL TBS‐T (0.05% Tween‐20) and once in TBS for 10 min, slides were incubated in the dark for 1hr at 20°C with 100μL of Goat Anti-Human IgG Fc (DyLight 650) pre-adsorbed secondary antibody (ab98622, Abcam, UK), using a working dilution of 1:5000 in blocking agent. The arrays were incubated for IgA detection with 100 μL of Goat Anti-Human IgA alpha chain (DyLight 650) pre-adsorbed secondary antibody (ab96998, Abcam, UK), using a working dilution of 1:5000 in blocking agent. Slides were rinsed in de-ionised water and dried by centrifugation at 200 x g for 2 min.
|
| Scan protocol |
Slides were scanned in an InnoScan 710 (Innopsys, France) with the photomultiplier tube set to 40% for 635 nm. Image analysis and data quantification was carried out using Mapix v9.1.0 - Microarray image acquisition and analysis software (Innopsys, France).
|
| Description |
PT_IgA_Slide_4.gpr
A historical cohort of individuals with laboratory-confirmed gonococcal infection
|
| Data processing |
Microarray spot intensities were quantified using Mapix v9.1.0 Microarray Image Analysis software (Innopsys, France), which utilises automatic background subtraction for each spot. The spot intensities for each protein were recorded in quintuplicate; arithmetic means were determined, and spot intensities for buffer-only controls were subtracted
|
| |
|
| Submission date |
Jun 12, 2024 |
| Last update date |
Aug 15, 2024 |
| Contact name |
Fidel Ramirez Bencomo |
| E-mail(s) |
[email protected]
|
| Organization name |
The University of Manchester
|
| Department |
FBMH/SBS/EIGen
|
| Lab |
JPD Lab
|
| Street address |
Michael Smith Building
|
| City |
Manchester |
| State/province |
England |
| ZIP/Postal code |
M13 9PT |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL34580 |
| Series (1) |
| GSE269648 |
Profiling IgG and IgA antibody responses during vaccination and infection in a population at high risk of gonorrhoea |
|
