Rabbit IgG negative control for ChIP

Product no: AS21 4694

AS21 4694  | Clonality:  Polyclonal  |  Host:  Rabbit  | Reactivity: Negative control IgG for chromatin immunoprecipiation assays (ChIP)

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  • Product Info
  • Host: Rabbit
    Clonality: Polyclonal
    Purity: Total IgG. Protein A purified in 2 mM phosphate, 30 mM NaCl, ph 7.8, 0.02% sodium azide. Contains sucrose for stabilization.
    Format: Liquid
    Quantity: 250 g at 1 g/l
    Storage: Store at 4C or -20C; 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: 200 (IF)
  • Application Examples
  • Chromatin immunoprecipitation using rabbit IgG negative control

    ChIP assay was done with rabbit polyclonal antibody against H3K4me3 (AS16 3190)  using chromatin from sheared 1 million HeLa cells. Rabbit IgG (AS21 4694) 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).

    Immunofluorescence using rabbit IgG negative control

    Detection of H3K4me3 on HeLa cells were stained with the rabbit polyclonal antibody against H3K4me3 (AS16 3190) (top) and with DAPI. Rabbit control IgG (AS21 4694) 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-H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in a blocking solution
    Secondary antibody: anti-rabbit antibody conjugated to Alexa488.
    Middle panel: staining of the nuclei with DAPI.

  • Background
  • Background: Rabbit 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 rabbit are used. This preparation contains a pool of the IgG subclasses from the serum of healthy rabbits.
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