Experiment Overview
| Repository ID: | FR-FCM-ZZU3 | Experiment name: | Figure.2.THP1 | MIFlowCyt score: | 97.75% |
| Primary researcher: | John Nash | PI/manager: | John Nash | Uploaded by: | John Nash |
| Experiment dates: | 2015-04-17 - 2015-04-18 | Dataset uploaded: | Oct 2016 | Last updated: | Jan 2017 |
| Keywords: | [THP1] [NFkB] [nuclear localization] | Manuscripts: | [28094900] |
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| Organizations: |
Stanford University, Stanford, CA (USA)
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| Purpose: | To perform proof-of-concept experiment for SLA-detection of nuclear localization of NF-kB in TNFa-stimulated THP1 myeloid cell lines. | ||||
| Conclusion: | NFkB nuclear import is successfully visualized using flow cytometry for THP-1 cell lines, with a 2-3 fold increase in signal upon stimulation. These results were substantiated by confocal microscopy experiments done in parallel, showing visually what SLA was able to quantify by flow cytometry. | ||||
| Comments: | This experiment, along with additional experiments using the U-937 monocytic cell line demonstrates proof-of-concept for the SLA method. | ||||
| Funding: | Cancer Biology Training Grant 2T32CA009302-36A1 | ||||
| Quality control: | Negative control: no primary antibody Positive control: double-strand DNA - Histone H3 interaction Experiment: NFkB - dsDNA interaction | ||||
Experiment variables
| Conditions | |
|---|---|
| · untreated | Sample_THP1_CD45_H3_1_002_Untreated_Duplicate 1_Ungated_CD45:H3_Untreated.fcs · Sample_THP1_CD45_H3_2_006_Untreated_Duplicate 2_Ungated_CD45:H3_Untreated.fcs · Sample_THP1_NFkB_dsDNA_1_004_Untreated_Duplicate 1_Ungated_NFkB:dsDNA_Untreated.fcs · Sample_THP1_NFkB_dsDNA_2_008_Untreated_Duplicate 2_Ungated_NFkB:dsDNA_Untreated.fcs · Sample_THP1_dsDNA_H3_1_003_Untreated_Duplicate 1_Ungated_dsDNA:H3_Untreated.fcs · Sample_THP1_dsDNA_H3_2_007_Untreated_Duplicate 2_Ungated_dsDNA:H3_Untreated.fcs |
| · TNFa | Sample_THP1_CD45_H3_1_002_TNFa_Duplicate 1_Ungated_CD45:H3_TNFa.fcs · Sample_THP1_CD45_H3_2_006_TNFa_Duplicate 2_Ungated_CD45:H3_TNFa.fcs · Sample_THP1_NFkB_dsDNA_1_004_TNFa_Duplicate 1_Ungated_NFkB:dsDNA_TNFa.fcs · Sample_THP1_NFkB_dsDNA_2_008_TNFa_Duplicate 2_Ungated_NFkB:dsDNA_TNFa.fcs · Sample_THP1_dsDNA_H3_1_003_TNFa_Duplicate 1_Ungated_dsDNA:H3_TNFa.fcs · Sample_THP1_dsDNA_H3_2_007_TNFa_Duplicate 2_Ungated_dsDNA:H3_TNFa.fcs |
| Individuals | |
|---|---|
| · CD45_H3 | Sample_THP1_CD45_H3_1_002_TNFa_Duplicate 1_Ungated_CD45:H3_TNFa.fcs · Sample_THP1_CD45_H3_1_002_Untreated_Duplicate 1_Ungated_CD45:H3_Untreated.fcs · Sample_THP1_CD45_H3_2_006_TNFa_Duplicate 2_Ungated_CD45:H3_TNFa.fcs · Sample_THP1_CD45_H3_2_006_Untreated_Duplicate 2_Ungated_CD45:H3_Untreated.fcs |
| · dsDNA_H3 | Sample_THP1_dsDNA_H3_1_003_TNFa_Duplicate 1_Ungated_dsDNA:H3_TNFa.fcs · Sample_THP1_dsDNA_H3_1_003_Untreated_Duplicate 1_Ungated_dsDNA:H3_Untreated.fcs · Sample_THP1_dsDNA_H3_2_007_TNFa_Duplicate 2_Ungated_dsDNA:H3_TNFa.fcs · Sample_THP1_dsDNA_H3_2_007_Untreated_Duplicate 2_Ungated_dsDNA:H3_Untreated.fcs |
| · NFkB_dsDNA | Sample_THP1_NFkB_dsDNA_1_004_TNFa_Duplicate 1_Ungated_NFkB:dsDNA_TNFa.fcs · Sample_THP1_NFkB_dsDNA_1_004_Untreated_Duplicate 1_Ungated_NFkB:dsDNA_Untreated.fcs · Sample_THP1_NFkB_dsDNA_2_008_TNFa_Duplicate 2_Ungated_NFkB:dsDNA_TNFa.fcs · Sample_THP1_NFkB_dsDNA_2_008_Untreated_Duplicate 2_Ungated_NFkB:dsDNA_Untreated.fcs |
| Sample Type | |
|---|---|
| · Duplicate1 | Sample_THP1_CD45_H3_1_002_TNFa_Duplicate 1_Ungated_CD45:H3_TNFa.fcs · Sample_THP1_CD45_H3_1_002_Untreated_Duplicate 1_Ungated_CD45:H3_Untreated.fcs · Sample_THP1_NFkB_dsDNA_1_004_TNFa_Duplicate 1_Ungated_NFkB:dsDNA_TNFa.fcs · Sample_THP1_NFkB_dsDNA_1_004_Untreated_Duplicate 1_Ungated_NFkB:dsDNA_Untreated.fcs · Sample_THP1_dsDNA_H3_1_003_TNFa_Duplicate 1_Ungated_dsDNA:H3_TNFa.fcs · Sample_THP1_dsDNA_H3_1_003_Untreated_Duplicate 1_Ungated_dsDNA:H3_Untreated.fcs |
| · Duplicate2 | Sample_THP1_CD45_H3_2_006_TNFa_Duplicate 2_Ungated_CD45:H3_TNFa.fcs · Sample_THP1_CD45_H3_2_006_Untreated_Duplicate 2_Ungated_CD45:H3_Untreated.fcs · Sample_THP1_NFkB_dsDNA_2_008_TNFa_Duplicate 2_Ungated_NFkB:dsDNA_TNFa.fcs · Sample_THP1_NFkB_dsDNA_2_008_Untreated_Duplicate 2_Ungated_NFkB:dsDNA_Untreated.fcs · Sample_THP1_dsDNA_H3_2_007_TNFa_Duplicate 2_Ungated_dsDNA:H3_TNFa.fcs · Sample_THP1_dsDNA_H3_2_007_Untreated_Duplicate 2_Ungated_dsDNA:H3_Untreated.fcs |
