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Sample GSM8010268 Query DataSets for GSM8010268
Status Public on Jan 10, 2024
Title SAMPLE 6 -Prime-boost vac regime Female RAM Mouse 32
Sample type protein
 
Source name Serum
Organism Mus musculus
Characteristics vaccine regimen: prime/boost
vaccine: RAM (H1N1)
time point: d14 dpb
mouse id: 32.0
Treatment protocol n/a
Growth protocol n/a
Extracted molecule protein
Extraction protocol n/a
Label anti-IgG/Q800
Label protocol n/a
 
Hybridization protocol Serum and BALF were diluted 1:100 in a protein array blocking buffer (GVS, Sanford, ME, USA), supplemented with E. coli lysate (GenScript, Piscataway, NJ, USA) to a final concentration of 10 mg/mL, and preincubated at room temperature (RT) for 30 min. Concurrently, arrays were rehydrated in blocking buffer (without lysate) for 30 min. Blocking buffer was removed, and arrays were probed with preincubated serum samples using sealed chambers to prevent cross-contamination of samples between the pads. Arrays were incubated overnight at 4  C with gentle agitation. They were then washed at RT three times with Tris-buffered saline (TBS) containing 0.05% Tween 20 (T-TBS), biotin-conjugated goat anti-mouse IgA and Biotin-conjugated anti-mouse IgG (Jackson Immuno Research Laboratories, Inc., West Grove, PA, USA) were diluted 1:400 in blocking buffer and applied to separate arrays for 1 h, RT with gentle agitation. Arrays were washed three times with T-TBS, followed by incubation with streptavidin-conjugated Qdot655 (Thermo Fisher Scientific, Waltham, MA, USA) diluted 1:200 in blocking buffer for 1 h, RT. Arrays were washed three times with T-TBS and once with water. Arrays were air dried by centrifugation at 500 g for 5 min. Images were acquired using the ArrayCAM imaging system from Grace Bio-Labs (Bend, OR, USA). Spot and background intensities were measured using an annotated grid (.gal) file. Mean fluorescence across antigens grouped by isotypes were used for subsequent analysis. The different antigens were acquired from Sino biological (Wayne, PA, USA).
Scan protocol Images were acquired using the ArrayCAM imaging system from Grace Bio-Labs (Bend, OR) with gain and exposure times of 50 and 400-1000ms, respectively.
Description Fig. 8 and 9
Data processing Spot and background intensities were measured using an annotated grid (.gal) file. Signal intensities (SI) for each antigen on the array were first background corrected by subtracting sample-specific T-PBS buffer signals from purified protein spot signals. For statistical analysis, A one-way ANOVA was performed. A P value below 0.05 was considered significan. All data analyses and graphs were performed using GraphPad Prism software version 9 (GraphPad Software Inc., San Diego, CA, USA).
 
Submission date Jan 10, 2024
Last update date Jan 10, 2024
Contact name Daniel Roberto Perez
E-mail(s) dperez1@uga.edu
Phone 7065425506
Organization name University of Georgia
Street address 953 College Station Road
City Athens
State/province GA
ZIP/Postal code 30602
Country USA
 
Platform ID GPL30424
Series (1)
GSE252919 FLUAV RAM-IGIP: A modified live influenza virus vaccine that enhances humoral and mucosal responses against influenza. Vaccine Encoding the IgA-Inducing Protein

Data table header descriptions
ID_REF
VALUE Signal intensity

Data table
ID_REF VALUE
210 48280
62 7049
224 37663
110 31982
122 24274
99 27029
205 13120
241 23491
173 496
73 6565
82 11498
221 9305
209 23713
53 35469
77 29075
88 13889
131 46593
239 54458
83 21477
124 39332

Total number of rows: 194

Table truncated, full table size 1 Kbytes.




Supplementary data files not provided
Processed data not provided for this record

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