Product Pathways - Companion Products
Normal Rabbit IgG #2729
| Applications | Reactivity | Sensitivity | Source |
|---|---|---|---|
| ChIP | Endogenous | Rabbit |
Applications Key:
ChIP=Chromatin IP
Reactivity Key:
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 2729:
- ChIP Agarose, ChIP Magnetic
Specificity / Sensitivity
Source / Purification
The purified antibody is not directed against any known antigen. It was isolated from naive rabbit sera and prepared by Protein A chromatography.
Chromatin IP
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 IP
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.
Description
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.
Directions for Use
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.
Background
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.
Application References
- Kong, D.K. et al. (2010) Mol Biol Cell 21, 1335-49. Applications: ChIP
- Shukla, S. et al. (2011) Nature 479, 74-9. Applications: ChIP
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 9002 SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
- 9003 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
- 9006 ChIP-Grade Protein G Magnetic Beads
- 9007 ChIP-Grade Protein G Agarose Beads
- 9725 Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb
- 9726 Di-Methyl-Histone H3 (Lys4) Antibody
- 9751 Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb
- 9753 Di-Methyl-Histone H3 (Lys9) Antibody
- 4069 Pan-Methyl-Histone H3 (Lys9) Antibody
- 9756 Tri-Methyl-Histone H3 (Lys27) Antibody
- 9649 Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb
- 9677 Acetyl-Histone H3 (Lys9/Lys14) Antibody
- 2629 Rpb1 CTD (4H8) Mouse mAb
For Research Use Only. Not For Use In Diagnostic Procedures.
