ANTI-SPO11 CLONE 180

General description

Meiotic recombination protein SPO11 (UniProt Q9WTK8) is encoded by the Spo11 gene (Gene ID 26972) in murine species. SPO11 is a widely conserved protein that is involved in meiotic recombination and mediates DNA cleavage that forms the double-strand breaks that initiate meiotic recombination. Active site of SPO11 contains a tyrosine, which ligates and dissociates with DNA to promote break formation. One SPO11 protein is involved per strand of DNA, hence, two SPO11 proteins are involved in each double stranded break event. High levels of SPO11 are found only in the testis where expression is restricted primarily to meiotic germ cells and it is not expressed in spermatogonia. Very low levels of SPO11 have been reported in thymus, brain, and oocytes of embryonic ovary, but it is not detected in adult ovary. Mouse and human SPO11 exhibit about 82% sequence homology. Ref.: Keeney, S., et al. (1997). Cell 375-384.

Specificity

Clone 180 detected SPO11alpha and SPO11beta in wild-type, but not Spo11-knockout mouse testes lysates by IP-Western blotting.

Immunogen

Recombinant mouse SPO11 with N-terminal S-tag and C-terminal His-tag.

Application

Anti-SPO11, clone 180, Cat. No. MABE1167, is a highly specific mouse monoclonal antibody that targets SPO11 and has been tested in and Immunocytochemistry, Immunoprecipitation, and Western Blotting.

Immunoprecipitation Analysis: 3 µg from a representative lot immunoprecipitated SPO11alpha and SPO11beta from wild-type, but not Spo11-knockout mouse testes lysates (Courtesy of the Keeney Lab, MSKCC, New York, U.S.A.).

Western Blotting Analysis: A representative lot detected SPO11alpha and SPO11beta in immunoprecipitate from wild-type, but not SPO-knockout, mouse testes using the same antibody (Courtesy of the Keeney Lab, MSKCC, New York, U.S.A.).

Immunocytochemistry Analysis: A representative lot immunostained SPO11 in meiotic prophase I leptotene cells by fluorescence immunocytochemistry staining of chromosome spreads from isolated mouse spermatocytes hypotonically swollen, fixed with 1% formaldehyde and permeabilized with 0.1% Triton X-100 (Alpatov, R., et al. (2014). Cell. 157(4):869-881).

Immunoprecipitation Analysis: A representative lot immunoprecipitated SPO11alpha and SPO11beta as well as SPO11-oligo complexes from mouse testes lysates. Much elevated and greatly reduced SPO11-oligo complexes were found in Atm-/- and Dmc-/- testes IP, respectively, when comparing to wild-type testes IP (Lange, J., et al. (2011). Nature. 479(7372):237-240).

Western Blotting Analysis: Representative lots detected SPO11alpha and SPO11beta in immunoprecipitates obtained with the same antibody from mouse testes. Much reduced SPO11 was found in Atm-/- and Dmc-/- testes IPs due to meiotic arrest at prophase I (Biswas, U., et al. (2013). PLoS Genet. 9(12):e1003985; Lange, J., et al. (2011). Nature. 479(7372):237-240).

Research Category
Epigenetics & Nuclear Function

Quality

Identity Confirmation by Isotyping Test.

Isotyping Analysis: The identity of this monoclonal antibody is confirmed by isotyping test to be mouse IgG2aκ.

Target description

~44/40 kDa (SPO11beta/SPO11alpha) observed. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified.

Purified mouse IgG2aκ 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.