- NBP2-59202 - Novus Biologicals NR3C1 antibody

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Supportive validation

Submitted by: Novus Biologicals (provider)
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Experimental details: Western Blot: GR/NR3C1 Antibody (C.15200010) [NBP2-59202] - Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10^6 Raji or Molt cells were analyzed by Western blot using the antibody against hGR. Figure B. Analysis of extracts from 300,000 HeLa cells with the antibody against hGR (1 ug/mL).

Supportive validation

Submitted by: Novus Biologicals (provider)
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Experimental details: Western Blot: GR/NR3C1 Antibody (C.15200010) [NBP2-59202] - Immunoprecipitation using the antibody against hGR. The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 uL IP reaction solution) using 5 ug of the antibody directed against hGR. The IP was followed by Western blot analysis.

Supportive validation

Submitted by: Novus Biologicals (provider)
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Experimental details: ELISA: GR/NR3C1 Antibody (C.15200010) [NBP2-59202] - Sandwich ELISA The specificity of the antibody directed against hGR was assessed by sandwich ELISA. Figure A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR. Figure B: ELISA results using the monoclonal antibody against hGR at 0.5 ug/mL. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR.

Supportive validation

Submitted by: Novus Biologicals (provider)
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Experimental details: Immunohistochemistry: GR/NR3C1 Antibody (C.15200010) [NBP2-59202] - Immunoreactivity of the monoclonal antibody against hGR in rat CA1 neurons of hippocampus. The antibody was used at 2.5 ug/mL.

Supportive validation

Submitted by: Novus Biologicals (provider)
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Experimental details: Chromatin Immunoprecipitation: GR/NR3C1 Antibody (C.15200010) [NBP2-59202] - ChIP assays were performed using HeLa cells, the monoclonal antibody directed against GR and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 ug of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR.

Supportive validation

Submitted by: Novus Biologicals (provider)
Main image
Experimental details: Chromatin Immunoprecipitation: GR/NR3C1 Antibody (C.15200010) [NBP2-59202] - ChIP assays were performed using HeLa cells, the monoclonal antibody directed against GR and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 ug of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR.

NBP2-59202