ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - ChIP Validated Antibody and Primer Set

General description

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ Acetyl-Histone H3 (Lys9) set includes the anti-acetyl-histone H3 (Lys9) antibody, a negative control antibody (normal rabbit serum), and qPCR primers which amplify a 166 base pair region within the promoter of the human GAPDH gene. The acetyl-histone H3 (Lys9) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of acetyl-histone H3 (Lys9) associated chromatin.

Specificity

Acetyl-Histone H3 (Lys9)

Immunogen

The acetyl-histone H3 (Lys9) antiserum is made against a peptide corresponding to amino acids 4-14 of yeast histone H3 acetylated on lysine 9.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated HeLa cells (1 X 10⁶ cell equivalents) was subjected to chromatin immunoprecipitation using 1 μL of either a normal rabbit antiserum or Anti-Acetyl-Histone H3 (Lys9) serum and the Magna ChIP A Kit (Cat. #17-610) (Figure 2). Successful immunoprecipitation of acetyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter or primers amplifying the promoter of human β-globin, which is transcriptionally inactive in HeLa cells. Percent Input relative to standard curves for each qPCR primer set are shown, with immunoprecipitated DNA from control serum shown as (-) and acetylhistone H3 serum as (+).
Please refer to the EZ-Magna A ChIP (Cat. #17-408) or EZ-ChIP (Cat. #17-371) protocol for experimental details.

Western Blot Analysis:
Acid-extracted proteins from normal HeLa cells (Lane 1) and HeLa cells treated with 5mM sodium butyrate for 24 hours (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-Acetyl-Histone H3 (Lys9) (1:5000). Proteins were visualized using a goat-anti rabbit secondary antibody conjugated to HRP and chemiluminescent detection system (Please see figures).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Chromatin Biology

This ChIPAb+ Acetyl-Histone H3 (Lys9) Serum -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Packaging

25 assays per kit, ~1μL per chromatin immunoprecipitation

Quality

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated HeLa cells (1 X 10⁶ cell equivalents) was subjected to chromatin immunoprecipitation using 1 μL of either a normal rabbit antiserum or Anti-Acetyl-Histone H3 (Lys9) serum and the Magna ChIP A (Cat. #17-610) Kit.
Successful immunoprecipitation of acetylhistone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter (Please see figures).
Please refer to the EZ-Magna A ChIP (Cat. #17-408) or EZ-ChIP (Cat. #17-371) protocol for experimental details.

Target description

17 kDa

Physical form

Anti-Acetyl-Histone H3 (Lys9) (rabbit polyclonal serum). One vial containing 25 μL of antiserum containing 0.05% sodium azide before the addition of glycerol to 30%.

Normal Rabbit Serum. One vial containing 25 uL antiserum containing 0.05% sodium azide.

Control Primers. One vial containing 75 μL of 5 μM of each primer specific for for human GAPDH.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG CGA

Storage and Stability

Stable for 1 year at -20°C from date of receipt

Analysis Note

Control
Included negative control antibody normal rabbit serum and control primers specific for human GAPDH promoter.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.