Experiment Overview
| Repository ID: | FR-FCM-ZZUS | Experiment name: | Figure.S1.U937 | MIFlowCyt score: | 95.75% |
| Primary researcher: | John Nash | PI/manager: | John Nash | Uploaded by: | John Nash |
| Experiment dates: | 2014-04-30 - 2014-04-30 | Dataset uploaded: | Oct 2016 | Last updated: | Jan 2017 |
| Keywords: | [monocyte] [TNF] [NFkB] | Manuscripts: | [28094900] |
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| Organizations: |
Stanford University, Stanford, CA (USA)
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| Purpose: | Proof of concept SLA experiment using nuclear import of NFkB in U937 monocytic cells upon stimulation with TNFa. | ||||
| Conclusion: | SLA detected nuclear import of NFkB in U937 cells after stimulation with TNFa. | ||||
| Comments: | None | ||||
| Funding: | Cancer Biology Training Grant 5T32CA9302-35 | ||||
| Quality control: | We had a negative control consisting of the SLA reagents but without the interacting primary antibody pair. This along with an interaction not expected to happen (CD45:Histone H3) provided us with a sense of background noise, and a baseline signal we could use for comparison. The positive control was an interaction expected to happen (dsDNA:Histone H3) to provide an upper limit for the strength of signal, which we could compare with our NFkB:dsDNA signal. | ||||
Experiment variables
| Conditions | |
|---|---|
| · unstimulated | Specimen_001_A5_A05_cells_M1R2 10:3:13 1:100_No primary antibody_unstimulated.fcs · Specimen_001_A6_A06_cells_M1R2 10:3:13 1:100_CD45 H130:Histone H3_unstimulated.fcs · Specimen_001_A7_A07_cells_M1R2 10:3:13 1:100_dsDNA:Histone H3_unstimulated.fcs · Specimen_001_A8_A08_cells_M1R2 10:3:13 1:100_NFkB ab16502 2ug:mL:dsDNA_unstimulated.fcs |
| · TNFa | Specimen_001_D5_D05_cells_M1R2 10:3:13 1:100_No primary antibody_TNFa 10ng:mL 15min.fcs · Specimen_001_D6_D06_cells_M1R2 10:3:13 1:100_CD45 H130:Histone H3_TNFa 10ng:mL 15min.fcs · Specimen_001_D7_D07_cells_M1R2 10:3:13 1:100_dsDNA:Histone H3_TNFa 10ng:mL 15min.fcs · Specimen_001_D8_D08_cells_M1R2 10:3:13 1:100_NFkB ab16502 2ug:mL:dsDNA_TNFa 10ng:mL 15min.fcs |
| Individuals | |
|---|---|
| · no primary antibody | Specimen_001_A5_A05_cells_M1R2 10:3:13 1:100_No primary antibody_unstimulated.fcs · Specimen_001_D5_D05_cells_M1R2 10:3:13 1:100_No primary antibody_TNFa 10ng:mL 15min.fcs |
| · CD45:Histone H3 | Specimen_001_A6_A06_cells_M1R2 10:3:13 1:100_CD45 H130:Histone H3_unstimulated.fcs · Specimen_001_D6_D06_cells_M1R2 10:3:13 1:100_CD45 H130:Histone H3_TNFa 10ng:mL 15min.fcs |
| · dsDNA:Histone H3 | Specimen_001_A7_A07_cells_M1R2 10:3:13 1:100_dsDNA:Histone H3_unstimulated.fcs · Specimen_001_D7_D07_cells_M1R2 10:3:13 1:100_dsDNA:Histone H3_TNFa 10ng:mL 15min.fcs |
| · NFkB:dsDNA | Specimen_001_A8_A08_cells_M1R2 10:3:13 1:100_NFkB ab16502 2ug:mL:dsDNA_unstimulated.fcs · Specimen_001_D8_D08_cells_M1R2 10:3:13 1:100_NFkB ab16502 2ug:mL:dsDNA_TNFa 10ng:mL 15min.fcs |
