Mouse IgG negative control for ChIP

AS21 4695 | Clonality: Monoclonal | Host: Mouse | Reactivity: Negative control IgG for chromatin immunoprecipiation assays (ChIP)

Product Information

Host Mouse

Clonality Monoclonal

Purity Total IgG. Protein A affinity purified in 5 mM phosphate, 75 mM NaCl, ph 7.8 with 0.06% sodium azide. Contains sucrose for stabilization.

Format Liquid

Quantity 100 µg at 1 µg/µl

Storage Store at 4°C or -20°C; and make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.

Tested applications Chromatin immunoprecipitation (ChIP), Immunofluorescence (IF)

Recommended dilution 1 μg (ChIP), 1: 500 (IF)

Reactivity

Application examples

Application examples Chromatin immunoprecipiation using mouas IgG negative control

ChIP assay was done with rabbit polyclonal antibody against H3K27ac using chromatin from sheared 1 million HeLa cells. Mouse IgG (AS21 4695) served as a negative IP control. Amount of antibody was 1 μg/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. The graph shows recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Immunofluorsescence using mouse IgG negative control

Detection of H3K27ac on HeLa cells were stained with the primary antibody agains H3K27ac (top) and with DAPI. Mouse control IgG (AS21 4695) was used as a negative control (bottom row).
Fixation: 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA.
Primary antibody: anti-H3K27ac or mouse IgG negative control antibody (left) diluted 1:500 in a blocking solution
Secondary antibody: anti-rabbit antibody conjugated to Alexa594.
Middle panel: staining of the nuclei with DAPI.

Additional information

Background

Background Mouse IgG negative control for ChIP is suitable for chromatin immunoprecipitation (ChIP) and has been validated for this assay as well as for MeDIP, IF and other experiments where primary antibodies made in a mouse are used. This preparation contains a pool of the IgG subclasses from the serum of healthy mice.

Product citations

background:	Mouse IgG negative control for ChIP is suitable for chromatin immunoprecipitation (ChIP) and has been validated for this assay as well as for MeDIP, IF and other experiments where primary antibodies made in a mouse are used. This preparation contains a pool of the IgG subclasses from the serum of healthy mice.
Picture (footer):	ChIP assay was done with rabbit polyclonal antibody against H3K27ac using chromatin from sheared 1 million HeLa cells. Mouse IgG (AS21 4695) served as a negative IP control. Amount of antibody was 1 μg/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. The graph shows recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Detection of H3K27ac on HeLa cells were stained with the primary antibody agains H3K27ac (top) and with DAPI. Mouse control IgG (AS21 4695) was used as a negative control (bottom row). Fixation: 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. Primary antibody: anti-H3K27ac or mouse IgG negative control antibody (left) diluted 1:500 in a blocking solution Secondary antibody: anti-rabbit antibody conjugated to Alexa594. Middle panel: staining of the nuclei with DAPI.
Clonality:	Monoclonal
Format:	Liquid
Host:	Mouse
Purity:	Total IgG. Protein A affinity purified in 5 mM phosphate, 75 mM NaCl, ph 7.8 with 0.06% sodium azide. Contains sucrose for stabilization.
Quantity:	100 µg at 1 µg/µl
recommended dilution:	1 μg (ChIP), 1: 500 (IF)
storage:	Store at 4°C or -20°C; and make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications:	Chromatin immunoprecipitation (ChIP), Immunofluorescence (IF)