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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 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.
Product Usage Information
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.
| Application | Dilution |
|---|---|
| CUT&Tag | 1:50 |
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.
参考图片
CUT&Tag was performed with HeLa cells and Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751 and Normal Rabbit IgG, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 6 (upper), including GAPDH (lower), a known target gene of H3K4me3 (see our ChIP-qPCR figure).
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.
CUT&Tag was performed with HeLa cells and Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751 or Normal Rabbit IgG, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across GAPDH, a known target gene of H3K4me3 (see our ChIP-qPCR figure).