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Analysis Note

ControlIncludes normal rabbit IgG and primers specific for human β-globin.

Application

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1.4 µg of either Normal Rabbit IgG or Anti-JMJD1C and the Magna ChIP^® A Kit (Cat. # 17-610). Successful immunoprecipitation of JMJD1C associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin as a positive locus, and β-actin promoter primers as a negative locus. (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
HeLa nuclear extract was probed with Anti-JMJD1C (1 µg/mL). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates JMJD1C (~284 kDa). (Figure 3).
Immunocytochemistry Analysis:
Representative lot data.
Confocal fluorescent analysis of HeLa cells using Anti-JMJD1C (Red). Actin filaments have been labeled with Alexa Fluor™ 488 dye -Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus. (Figure 4).

This ChIPAb+ JMJD1C -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Research CategoryEpigenetics & Nuclear Function

Research Sub CategoryHistone Modifying Proteins

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.

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+JMJD1C set includes the JMJD1C antibody, a Normal Rabbit IgG, and control primers which amplify a 110 bp region of ChIP Primers, human β-globin. The JMJD1C and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of JMJD1C -associated chromatin.

JMJD1C (jumonji domain-containing protein 1C) is also known as TRIP8 and mutations in this histone demethylase have been linked to certain forms of autism. JMJD1C is also thought to work in reactivating genes in embryonic stem cells, pancreatic islet and gastric cancer. JMJD1C has also been implicated as a gene expression regulator. JMJD1C with WHISTLE have recently been shown to also play a role in mouse testis development. JMJD1C has been identified as a nuclear-receptor coactivator and is a candidate gene for autism.

Immunogen

KLH-conjugated linear peptide corresponding to human JMJD1C.

Legal Information

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

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

Other Notes

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

Packaging

25 assays per set. Recommended use: 1.4 µg (2 µL) of antibody per chromatin immuno-precipitation (dependent upon biological context).

Physical form

Affinity purified

Anti-JMJD1C (rabbit polyclonal). One vial containing 50 µL of purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl and 0.05% sodium azide before the addition of glycerol to 30%. Store at -20°C.
Concentration: 0.7 mg/mL
Normal Rabbit IgG. One vial containing 125 µg of Rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C.
ChIP Primers, human β-globin. One vial containing 75 µL of each primer (5 µM) specific for the human β-globin
promoter. Store at -20°C.
FOR: AGG ACA GGT ACG GCT GTC ATC
REV: TTT ATG CCC AGC CCT GGC TC

Quality

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1.4 µg of either Normal Rabbit IgG, or Anti-JMJD1C and the Magna ChIP^® A Kit (Cat. # 17-610).
Successful immunoprecipitation of JMJD1C associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin (Figure 1).
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

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.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Target description

~284 kDa observed