The negative control IgG from mouse has been extensively validated in chromatin immunoprecipitation (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy mice. This IgG preparation is intended for use as a negative control in ChIP, MeDIP, IF and other experiments performed with specific antibodies made in mouse. The negative control IgG from mouse should be used in parallel with the specific antibody at the same concentration. It is also included in many of our ChIP and MeDIP kits.
Mouse IgG (sample size)
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The negative Ctrl IgG from mouse has been extensively validated in chromatin profiling assays, like ChIP or CUT&Tag. It contains a spectrum of the IgG subclasses present in serum of healthy animals. This IgG preparation is intended for use as a negative control in ChIP or CUT&Tag experiments (but also in MeDIP, IF and other experiments) for specific antibodies made in mouse. The negative Ctrl IgG from mouse should be used for ChIP or CUT&Tag in parallel with specific antibody at the same concentration as the specific antibody.
| Lot | NA |
|---|---|
| Concentration | 1.0 µg/µl |
| Species reactivity | none |
| Type | Polyclonal, ChIP-grade, CUT&Tag-grade |
| Purity | Purified by protein A chromatography |
| Host | Mouse |
| Storage Buffer | 2 mM phosphate, 30 mM NaCl, ph 7.8; 0.04% sodium azide. Contains sucrose for stabilization |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
- Validation data
Figure 1. ChIP with the mouse IgG negative control antibody
ChIP assays were performed on sheared chromatin from 1 million HeLa cells using the mouse monoclonal antibody against H3K27ac (cat. No. C15200184) and the Hologic Diagenode ChIP kit. Mouse IgG (cat. No. C15400001) was used as a negative IP control. One microgram (µg) of each antibody was used per ChIP experiment. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. CUT&Tag results obtained with the rabbit IgG negative control antibody
CUT&Tag was performed on 50,000 K562 cells using 0.5 µg of the rabbit monoclonal antibody against H3K27me3 (cat. No. C15200181) and the Universal CUT&Tag kit (cat. No. C01070024). Mouse IgG (cat. No. C15400001) was used as a negative control. The resulting libraries were analyzed on a Fragment Analyzer. Figure 2 shows the results with the H3K27me3 antibody on top and the IgG negative control at the bottom.
Figure 3. Immunofluorescence with the mouse IgG negative control antibody
HeLa cells were stained with the mouse monoclonal antibody against H3K27ac (cat. No. C15200184) (top) and with DAPI. Mouse IgG (cat. No. C15400001) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with either the H3K27ac antibody or the mouse IgG negative control antibody (left), each diluted 1:500 in blocking solution; followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. - Publications
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