|
| Status |
Public on Jan 10, 2024 |
| Title |
TB_9611_Pre-BALF IgA |
| Sample type |
protein |
| |
|
| Source name |
Pre-BALF
|
| Organism |
Macaca fascicularis |
| Characteristics |
infection states: Symptomatic uncontrolled Mtb infection/TB disease
|
| Treatment protocol |
BALF dilted (1:8) or plasma diluted (1:100)
|
| Growth protocol |
N/A
|
| Extracted molecule |
protein |
| Extraction protocol |
Plasma was obtained from peripheral blood samples and the BAL procedure was performed as previously described via direct visualization of the airway and lavage of ~ 40 ml normal saline (0.9%). Animal protocols and procedures were approved by the University of Pittsburgh’s Institutional Animal Care and Use Committee before the collection of samples.
|
| Label |
Cy3 Goat Anti-Human Serum IgA
|
| Label protocol |
Cy3 Goat Anti-Human Serum IgA (Jackson ImmunoResearch Labs Cat# 109-165-011, RRID:AB 2337721)
|
| |
|
| Hybridization protocol |
Slides with 8 arrays on each were blocked with 3% BSA (GEMINi Bio-Products, standard grade) in PBS at 4°C overnight, washed with 0.1 % PBS-T, and then incubated with BALF (1:8) or plasma (1:100), diluted in 0.1% PBST with 3% BSA, for 4 hrs at 37°C. After washing, bound antibodies were detected by Cy3 Goat Anti-Human Serum IgA (Jackson ImmunoResearch Labs Cat# 109-165-011, RRID:AB 2337721) and Alexa 647 Goat Anti-Human IgG (Jackson ImmunoResearch Labs Cat# 109-605-008, RRID:AB_2337882. Slides were then washed with PBST, spun dry in a centrifuge, and scanned.
|
| Scan protocol |
The slides were scanned using the GenePix 4000 Microarray scanner system. Scanned images were analyzed with by the image-processing software G\enePix Pro 7.3.0.0 to analyze the images and measure the median pixel intensity (MPI).
|
| Description |
IgA
|
| Data processing |
The median pixel intensity of each feature was background subtracted. A minimum value of pixel intensity (400) was determined by the median pixel intensity of the spots with low quality, which was determined by the software quality control score for each spot depending on signal-to-noise ratio and spot shape. The scanned images were visually inspected. The spots with obvious local artifacts were excluded from analysis. The array features were printed in triplicate. The final data were averaged from the triplicates.
|
| |
|
| Submission date |
Sep 20, 2023 |
| Last update date |
Jan 10, 2024 |
| Contact name |
Jacqueline M. Achkar |
| E-mail(s) |
Jacqueline.achkar@einsteinmed.edu
|
| Phone |
7184302971
|
| Organization name |
Albert Einstein College of Medicine
|
| Street address |
1300 Morris Park Ave
|
| City |
Bronx |
| State/province |
NY |
| ZIP/Postal code |
10461 |
| Country |
USA |
| |
|
| Platform ID |
GPL30414 |
| Series (2) |
| GSE243658 |
Mucosal and systemic antigen-specific antibody responses correlate with protection against active tuberculosis in nonhuman primates (LAM63 IgA) |
| GSE243663 |
Mucosal and systemic antigen-specific antibody responses correlate with protection against active tuberculosis in nonhuman primates |
|
