Analysis Note

ControlHeLa nuclear extract
Included Positive Antigen Control: Catalog # 12-309, HeLa nuclear extract. Add an equal volume of Laemmli reducing sample buffer to 10 µL of extract and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced extract per lane for minigels.

Application

Research Sub CategoryTranscription Factors

Anti-REST Antibody is a rabbit polyclonal antibody for detection of REST also known as RE1-silencing transcription factor, X2 box repressor, neuron restrictive silencer factor & has been validated in WB.

Research CategoryEpigenetics & Nuclear Function

Updated information-Rest is glycosylated. From http://amsdottorato.cib.unibo.it/687/
From the summary:
'Finally, the glycosylation pattern of the REST protein was analysed, moving from the observation that the molecular weight calculated on REST sequence is about 116 kDa but using western blotting this transcription factor appears to have distinct apparent molecular weight (see Table 1.1): this difference could be explained by post-translational modifications of the proteins, like glycosylation. In fact recently, several studies underlined the importance of O-glycosylation in modulating transcriptional silencing, protein phosphorylation, protein degradation by proteasome and protein–protein interactions [Julenius et al., 2005; Zachara and Hart, 2006]. Deglycosilating analysis showed that REST protein in SH-SY5Y and HEK293 cells is Oglycosylated and not N-glycosylated. Moreover, using several combination of deglycosilating enzymes it is possible to hypothesize the presence of Gal-β(1-3)-GalNAc residues on the endogenous REST, while β(1-4)-linked galactose residues may be present on recombinant REST protein expressed in HEK293 cells. However, the O-glycosylation process produces an immense multiplicity of chemical structures and monosaccharides must be sequentially hydrolyzed by a series of exoglycosidase. Further experiments are needed to characterize all the post-translational modification of the transcription factor REST.' In particular the authors show that REST migrates at 200kd in SH-SY5Y cells.

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

NRSF (neuron-restrictive silencer factor, also designated XBR and REST for RE1-silencing factor) is a silencer protein that binds the DNA sequence element NRSE (neuron-restrictive silencer element). The binding of NRSF to the NRSE represses neuronal gene transcription in non-neuronal cells. Although NRSF is most highly expressed in non-neural tissues, it is also expressed in developing neurons and at low levels in the brain. NRSF contains nine zinc finger domains, but also exists as a C-terminally truncated form produced by alternative splicing. This variant, REST4, contains five of the zinc-finger domains and weakly binds DNA, yet is transported to the nucleus. NRSF associates with mSin3 and HDAC in ventricular myocytes, suggesting a role for NRSF outside the nervous system. Down-regulation of NRSF, which normally occurs upon neural differentiation, is necessary for the proper development of certain classes of neurons. NRSF is required to repress neuronal gene expression in vivo, in both extra-neural and undifferentiated neural tissue.

Immunogen

GST fusion protein corresponding to residues 801-1097 of human REST (repressor element RE-1 silencing transcription factor). Also known as NRSF (neuronal restricted silencing factor).

Legal Information

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.

Physical form

Protein A purified

Purified rabbit IgG in buffer containing 0.014 M phosphate buffer, pH 7.6, 0.175 M NaCl, 0.07% sodium azide, and 30% glycerol. Store at -20°C.

Format: Purified

Quality

Routinely evaluated by western blot with HeLa and Raji RIPA lysate.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected REST in nuclear extracts from HeLa and Raji cells.

Specificity

Recognizes REST, Mr 200 kDa

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, 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

120-200 kDa