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AccessionNameType
P-AGIL-2
hybridization
Agilent One-Color Microarray-Based Gene Expression (Hybridization) Version: 5.7 Agilent publication number: G4140-90040 URL: http://www.chem.agilent.com/Library/usermanuals/Public/G4140-90040_One-Color_GE_5.7.pdf This document describes Agilent's recommended procedures for the preparation and labeling of complex biological targets and hybridization, washing, scanning, and feature extraction of Agilent 60-mer oligonucleotide microarrays for microarray-based one-color gene expression analysis.
P-AGIL-28
labeling
P-AGIL-28
P-MTAB-32155
normalization data transformation protocol
The data were normalized using the percentile shift method to the 80th percentile.
P-MTAB-32154
array scanning protocol
Agilent SurePrint G3 mouse GE 8x60k microarray chips were scanned with an Agilent C Scanner with Surescan High Resolution Technology.
(Parameters: Scanning hardware = OTHER: Agilent C scanner, Scanning software = Scanning software)
P-MTAB-32160
nucleic acid extraction protocol
Cell pellets were resuspend in Trizol and frozen at -80oC. RNA was then extracted from the thawed samples via the phenol chloroform method.
(Parameters: Extracted product = total_RNA, Amplification = none)
P-MTAB-32156
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom- uninfected Kupffer cells.
P-MTAB-32150
growth protocol
3 C57BL/6 mice were injected with 200uL RPMI intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ Kupffer cells.
P-MTAB-32146
growth protocol
3 C57BL/6 mice were injected with 200uL RPMI intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ Kupffer cells.
P-MTAB-32152
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom+ infected Kupffer cells.
P-MTAB-32141
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom- uninfected Kupffer cells.
P-MTAB-32153
growth protocol
3 C57BL/6 mice were injected with 200uL RPMI intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ Kupffer cells.
P-MTAB-32140
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom- uninfected Kupffer cells.
P-MTAB-32144
growth protocol
3 C57BL/6 mice were injected with 200uL RPMI intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ Kupffer cells.
P-MTAB-32147
growth protocol
3 C57BL/6 mice were injected with 200uL RPMI intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ Kupffer cells.
P-MTAB-32158
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom- uninfected Kupffer cells.
P-MTAB-32148
growth protocol
3 C57BL/6 mice were injected with 200uL RPMI intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ Kupffer cells.
P-MTAB-32142
growth protocol
3 C57BL/6 mice were injected with 200uL RPMI intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ Kupffer cells.
P-MTAB-32159
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom- uninfected Kupffer cells.
P-MTAB-32163
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom+ infected Kupffer cells.
P-MTAB-32162
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom+ infected Kupffer cells.
P-MTAB-32143
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom- uninfected Kupffer cells.
P-MTAB-32157
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 12hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom- uninfected Kupffer cells.
P-MTAB-32139
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 12Hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom+ infected Kupffer cells.
P-MTAB-32149
growth protocol
3 C57BL/6 mice were injected with 200uL RPMI intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ Kupffer cells.
P-MTAB-32151
growth protocol
4 C57BL/6 mice were injected with 3x10e7 TdTom.LV9 amastigotes intravenously. 2hr later, the livers were removed, pooled and digested in with 0.4mg/mL warmed liberase TL (Roche) at 37oC for 45min, then homogenised. Digested livers were passed over 100uM cell strainers, washed in PBS containing 2%FCS and passed over a 33% percoll density gradient. The remaining cell pellet was washed, RBC lysed and the remaining cells labelled with F4/80, GR-1 and anti-CRIg and sorted on a MoFlo (DAKO) fluorescence activated cell sorter to isolate CRIg+ F4/80+ TdTom+ infected Kupffer cells.