17-641

ChIPAb+ REST - ChIP Validated Antibody and Primer Set

• UNSPSC Code: 12352203
• eCl@ss: 32160702
• NACRES: NA.52

Properties

• antibody form: affinity purified immunoglobulin
• Quality Level: 100
• clone: polyclonal
• species reactivity: rat, human, rabbit, mouse
• technique(s): ChIP: suitable; western blot: suitable
• UniProt accession no.: Q13127

Description

General description

FUNCTION:Transcriptional repressor which binds neuron-restrictive silencer element (NRSE) and represses neuronal gene transcription in non-neuronal cells. Restricts the expression of neuronal genes by associating with two distinct corepressors, mSin3 and CoREST, which in turn recruit histone deacetylase to the promoters of REST-regulated genes. Mediates repression by recruiting the BHC complex at RE1/NRSE sites which acts by deacetylating and demethylating specific sites on histones, thereby acting as a chromatin modifier.

SUBUNIT STRUCTURE: Interacts with SIN3A, SIN3B and RCOR1.

SUBCELLULAR LOCATION: NucleusProbable.

TISSUE SPECIFICITY: Ubiquitous. Expressed at higher levels in the tissues of the lymphocytic compartment, including spleen, thymus, peripheral blood lymphocytes and ovary.

SEQUENCE SIMILARITY:Contains 9 C2H2-type zinc fingers.

SEQUENCE CAUTION: The sequence AAA98503.1 differs from that shown. Reason: Frameshift at positions 189, 212 and 1087.

The sequence AAH38985.1 differs from that shown. Reason: Miscellaneous discrepancy. Contaminating sequence. Potential poly-A sequence.

The REST purified antibody is made against a GST fusion protein corresponding to amino acids 801-1097 of human REST.

Specificity

Recognizes REST, Mr 120-200 kDa.

Application

Epitope: a.a. 801-1097

Packaging

25 assays per set. Recommended use: ~2 μg antibody per chromatin immunoprecipitation (dependent upon biological context).

Quality

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (4 x 10⁶ cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal rabbit IgG or Anti-REST antibody and the Magna ChIP A Kit (Part No. 17-610) (Please see figures). Successful immunoprecipitation of REST-associated DNA fragments was verified by qPCR using Control Primers.

Physical properties

120 kDa predicted, 200 kDa observed

Physical form

Anti-REST (rabbit polyclonal IgG). One vial containing 50 μg of affinity purified antibody in 50 μL PBS with 0.07% sodium azide, and 30% glycerol. Store at -20°C.

Normal Rabbit IgG. One vial containing 125 μg purified rabbit IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

ChIP Primers, Synapsin RE1. One vial containing 75 μL of 5 μM of each primer specific for the RE1 site of human Synapsin. Store at -20°C.
FOR: GGT GCT GAA GCT GGC AGT
REV: TGG GTT TTA GGA CCA GGA TG

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.

Other Notes

Human, mouse, rat. Not tested in other species.

Includes negative control rabbit IgG antibody and control primers specific for the RE1 site of human Synapsin.

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.

Safety Information

• Storage Class: 10 - Combustible liquids