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Sample GSM3474025 Query DataSets for GSM3474025
Status Public on Dec 19, 2018
Title ZyA + β2 ADR
Sample type protein
 
Source name Peritoneal monocytes/macrophages
Organism Mus musculus
Characteristics strain: C57BL/6
cell type: peritoneal macrophages
Extracted molecule protein
Extraction protocol 1. Cells were washed 5 times with ice cold PBS and remaining PBS removed
2. 100 µl of Full Moon Lysis Buffer was added and cells detached using a cell scraper and transferred to a microcentrifuge tube. One tube of Lysis Beads was added.
3. Mixture were vortexed for 60 seconds and incubated on ice for 10 minutes. Procedure was repeated 5 times.
4. Mixture was centrifuged at 10000 g for 5 minutes at 4°C.
5. Liquid was transferred to a fresh tube and centrifuged at 18000 g for 20 minutes at 4°C.
6. Supernatant was transferred to a fresh tube.
Label Cy3
Label protocol 1. 100 µg of protein was used and Labeling Buffer was added to bring the volume to 75 µl.
2. 3 µl of Biotin/DMF solution was added and the mixture incubated at room temperature for 2 hours with vortexing every 10 minutes.
3. 35 µl of Stop Reagent was added and solution mixed by vortexing. Incubation for 30 minutes.
 
Hybridization protocol 1. 6 ml Protein Coupling Solution and one tube of biotinylated sample was mixed (Protein Coupling Mix)
2. Slides were incubated in Protein Coupling Mix for 2 hours on an orbital shaker (35 rpm)
3. For Detection, 60 µl of Cy3-streptavidin (0.5 mg/ml) were added to 60 ml of Detection Buffer.
4. Slides were submerged in 30 ml of Cy3-streptavidin solution for 20 minutes on an orbital shaker (35 rpm) at room temperature in the dark.
Scan protocol Scans were performed by Full Moon BioSystems
Description Peritoneal monocytes/macrophages from C57BL/6 mice treated with Zymosan A and β2 ADR agonist were used following 12 h of ZyA-induced peritonitis. Protein and phosphorylation (Phospho Explorer Antibody Array, FullMoonBioscience, #PEX100) profiling of peritoneal monocytes (pooled lavages from 3-4 mice / condition) was carried out according to the manufacturer’s instructions.
Data processing Ratio data file is available on the series record and includes the following columns:
AB: Antibody Name
ID: UniProt ID
PER1: Protein Expression Ratio (ZyA + RGM-A/ZyA)
PER2: Protein Expression Ratio (ZyA + RGM-A + β2 ADR/ZyA)
PER3: Protein Expression Ratio (ZyA + RGM-A + β2 ADR/ZyA + RGM-A)
PA: Phosphorylation Antibody
PS: Phosphorylation Site
PPER1: Phospho Protein Expression Ratio (ZyA + RGM-A/ZyA)
PPER2: Phospho Protein Expression Ratio (ZyA + RGM-A + β2 ADR/ZyA)
PPER3: Phospho Protein Expression Ratio (ZyA + RGM-A + β2 ADR/ZyA + RGM-A)
 
Submission date Nov 15, 2018
Last update date Dec 19, 2018
Contact name Andreas Körner
Organization name University of Tuebingen
Department Clinic of Anesthesiology and Intensive Care Medicine
Street address Hoppe-Seyler-Straße 3
City Tuebingen
ZIP/Postal code 72076
Country Germany
 
Platform ID GPL18873
Series (1)
GSE122569 Role of Sympathic Nervous System and RGM-A in the Resolution of Inflammation

Supplementary file Size Download File type/resource
GSM3474025_4000020542.txt.gz 119.9 Kb (ftp)(http) TXT
Processed data are available on Series record

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