Immunoprecipitation of 4E-BP2 from C2C12 cell extracts using Normal Rabbit IgG #2729 (lane 2) or 4E-BP2 Antibody #2845 (lane 3). Lane 1 is 10% input. Western blot analysis was performed using 4E-BP2 Antibody #2845.
Chromatin immunoprecipitations were performed using digested chromatin from HeLa cells and either Histone H3 (D2B12) XP® Rabbit mAb (ChIP Formulated) #4620 (lane 2), Rpb1 CTD (4H8) Mouse mAb #2629 (lane 3), Di-Methyl Histone H3 (Lys9) Antibody #9753 (lane 4) or Normal Rabbit IgG #2729 (lane 5). Purified DNA was analyzed by standard PCR methods using SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human a Satellite Repeat Primers #4486. PCR products were observed for each primer set in the input sample (lane 1) and various protein-specific immunoprecipitations but no PCR products were observed with immunoprecipitation using Normal Rabbit IgG #2729 (lane 5).
Chromatin immunoprecipitations were performed using digested chromatin from HeLa cells and the indicated antibodies. Purified DNA was analyzed by quantitative real-time PCR, using SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human α Satellite Repeat Primers #4486. The relative abundance of each DNA sequence enriched by Normal Rabbit IgG (red) is compared to the amount of the same DNA sequence enriched by the protein-specific immunoprecipitations.
Normal Rabbit IgG is an unconjugated rabbit polyclonal antibody that is routinely used as a non-specific IgG control in chromatin immunoprecipitation using our SimpleChIP® Enzymatic Chromatin IP Kits #9002 and #9003.
Important! Dilute this control antibody to the same concentration (not dilution) as the specific test antibody in the chromatin immunoprecipitation. This is typically 1 μl (1 μg) to 5 μl (5 μg) of control antibody for one immunoprecipitation. Higher background signal may result if an excessive amount of rabbit IgG isotype control is used.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
The purified antibody is not directed against any known antigen. It was isolated from naive rabbit sera and prepared by Protein A chromatography.
Normal Rabbit IgG is an isotype control antibody, which is used to estimate the non-specific binding of target primary antibodies due to Fc receptor binding or other protein-protein interactions. An isotype control antibody should have the same immunoglobulin type and be used at the same concentration as the test antibody.
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