ANTI-ACETYL SMC3 (LYS105/106) CLONE 21A7

General description

Structural maintenance of chromosomes protein 3 (UniProt Q9UQE7; also known as Bamacan, Basement membrane-associated chondroitin proteoglycan, Chondroitin sulfate proteoglycan 6, Chromosome-associated polypeptide, hCAP, SMC protein 3, SMC-3) is encoded by the SMC3 (also known as BAM, BMH, CDLS3, CSPG6, SMC3L1) gene (Gene ID 9126) in human. Cohesion is a nuclear tripartite complex composed of Rad21 and two structural maintenance of chromosomes proteins, SMC1 and SMC3. Cohesin plays an important role in sister chromatid cohesion, as well as in DNA repair and gene expression regulation. To establish sister chromatid cohesion during S phase, SMC3 is acetylated (on K105 and K106 in human) by two cohesin acetyltransferases (CoATs), ESCO1 and ESCO2, that differ in their N-terminal domains and expression during development and across the cell cycle. ChIP-seq analysis reveals that ESCO1 is recruited by cohesin to over 11,000 chromatin sites occupied by cohesin and CTCF that mediate long-range chromatin interactions and regulate transcription globally. On ther other hand, ESCO2 is infrequently enriched at sites targeted by REST/NRSF (RE1-silencing transcription factor/neuron-restrictive silencer factor) that represses transcription of neuron-specific genes. SMC3 acetylation is thought to induce conformational changes of the dimeric hinge structure during S phase and to allow dimer opening and loading of DNA.

Specificity

Clone 21A7 bound Lys105/106-diacetylated SMC3 peptide with 2-fold higher affinity than Lys105-monoacetylated SMC3 peptide (Kd = 0.04 and 0.09 µg/mL, respectively), while displaying much reduced affinity toward Lys105-monoacetylated (Kd = 0.73 µg/mL) and little affinity toward nonacetylated (Kd = 2.99 µg/mL) peptide (Rahman, S., et al. (2015). Proc. Natl. Acad. Sci. U. S. A. 112(36):11270-11275).

Immunogen

KLH-conjugated linear peptide corresponding to the target region sequence of human/mouse/rat SMC3 containing acetylated Lys105 and Lys106.

Application

Anti-acetyl SMC3 (Lys105/106), clone 21A7, Cat. No. MABE1073, is a highly specific mouse monoclonal antibody that targets SMC3 Lys105/106 acetylation and has been tested in Chromatin Immunoprecipitation (ChIP), ChIP-seq, ELISA, and Western Blotting.

ELISA Analysis: A representative lot bound Lys105/106-diacetylated SMC3 peptide with 2-fold higher affinity than Lys105-monoacetylated SMC3 peptide (Kd = 0.04 and 0.09 µg/mL, respectively), while displaying much reduced affinity toward Lys105-monoacetylated (Kd = 0.73 µg/mL) and little affinity toward nonacetylated (Kd = 2.99 µg/mL) peptide (Rahman, S., et al. (2015). Proc. Natl. Acad. Sci. U. S. A. 112(36):11270-11275).

Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected enhanced SMC3 occupancy at target chromatin sites in HDAC8-null HCT116 cells than wild-type HCT-116 cells (Rahman, S., et al. (2015). Proc. Natl. Acad. Sci. U. S. A. 112(36):11270-11275).

ChIP-seq Analysis: A representative lot detected SMC3 chromosome 11 enrichment sites, including 81% of Rad21 target sites and 70% of Esco1 target sites, by ChIP-seq (Rahman, S., et al. (2015). Proc. Natl. Acad. Sci. U. S. A. 112(36):11270-11275).

Western Blotting Analysis: A representative lot detected SMC3 Lys105/106 acetylation in HeLa cells. ESCO1 or ESCO2 depletion by shRNA treatment reduced SMC3 Lys105/106 acetylation level, while dual ESCO1/2 depletion resulted in most profound SMC3 Lys105/106 acetylation reduction (Rahman, S., et al. (2015). Proc. Natl. Acad. Sci. U. S. A. 112(36):11270-11275).

Research Category
Epigenetics & Nuclear Function

Quality

Evaluated by Western Blotting in human retinal epithelial pigment (REP) cells.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected a reduced SMC3 Lys105/106 acetylation in 10 µg of lysate from N-acetyltransferase ESCO1-knockout human retinal epithelial pigment (REP) cells than in lysate from wild-type REP cells.

Target description

~160 kDa observed. 142.0/141.5/141.6/138.4/140.7 kDa (bovine/human/mouse/rat/xenopus) calculated. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified.

Purified mouse IgG2b in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

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.