ChIPAb+ CBX8 - 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+ CBX8 set includes a CBX8 antibody, a Normal Rabbit IgG, and positive control primers which amplify 138 bp of the human HOXA2 promoter region. The CBX8 antibody and negative control IgG are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of CBX8 associated chromatin.

The family of Heterochromatin Proteins ("Chromobox Homolog", CBX) are highly conserved adapter molecules, which have important functions in the cell nucleus. These functions include gene repression by heterochromatin formation, transcriptional activation, regulation of binding of cohesion complexes to centromere, sequesteration of genes to nuclear periphery, transcriptional arrest, maintenance of heterochromatin integrity, gene repression at single nucleosome level and gene repression by heterochromatization of euchromatin.

Specificity

Wide species reactivity expected based upon conservation of immunogen sequence

Immunogen

Epitope: Human CBX8 (a.a. 350--389)

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from Ntera-2 cells fixed for 15 min (1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-CBX8 (Part No. CS207348) and the Magna ChIP HiSens (Cat. # 17-10460). Successful immunoprecipitation of CBX8 associated DNA fragments was verified by qPCR using ChIP Primers, HoxA2 upstream (Part No. CS203200) as a positive locus, and GAPDH promoter primers (Part No. 22-004) as a negative locus. Data are presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the Magna ChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens(Cat. # 17-10461) protocol for experimental details.

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from Ntera-2 cells fixed for 15 min ( 1e5 and 1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-CBX8 (Part No. CS207348) and the Magna ChIP HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of CBX8 associated DNA fragments was verified by qPCR using ChIP Primers, Hox A2 upstream (Part No. CS203200).
Please refer to the MagnaChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens (Cat. # 17-10461) protocol for experimental details.

Western Blotting Analysis: 0.1 µg/mL of this antibody detected CBX8 on 5, 15 and 50 µg of HeLa whole cell lysate and 50 µg of 293T whole cell lysate.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Chromatin Biology

Use ChIPAb+ CBX8 ChIP Validated rabbit polyclonal antibody for the detection of CBX8 also known as Chromobox protein homolog 8 & Polycomb 3 homolog.

Packaging

25 assays per set. Recommended use: 2.0 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).

Quality

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from Ntera-2 cells fixed for 15 min (1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-CBX8 (Part No. CS207348) and the Magna ChIP^® HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of CBX8 associated DNA fragments was verified by qPCR using ChIP Primers, HoxA2 upstream (Part No. CS203200).
Please refer to the MagnaChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens (Cat. # 17-10461) protocol for experimental details.

Target description

~43kDa

Physical form

Affinity purified

Anti-CBX8 (Rabbit Polyclonal), Part No. CS207348. One vial containing 50 µg affinity purified rabbit polyclonal in 72 uL buffer containing Tris-citrate/phosphate buffer (pH 7 to 8) and 0.09% sodium azide, before the addition of glycerol to 30% (0.7 mg/ml final). Store at -20°C.

Normal Rabbit IgG, Part No. PP64B. One vial containing 125 μg of purified Rabbit IgG in 125 μL storage buffer containing 0.05% sodium azide. Store at -20°C.

ChIP Primers, HOXA2 Upstream. Part No. CS203200. One vial containing 75 μL of 5 μM of each primer specific for human HOXA2 promoter region (chr7:27142948-27143085, hg19 build). Store at -20°C.
FOR: AGG AAA GAT TTT GGT TGG GAA G
REV: AAA AAG AGG GAA AGG GAC AGA C

Format: Purified

Storage and Stability

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance

Analysis Note

Control
Includes normal rabbit IgG and primers specific for the human HOXA2 promoter region.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

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.