Anti-RLIM/Rnf12 Antibody (C15-1317-000)

General description

E3 ubiquitin-protein ligase RLIM (UniProt Q9WTV7; also known as Hoxa1 regulated, LIM domain-interacting RING finger protein, R-LIM, RING finger LIM domain-binding protein, Ring finger protein 12) is encoded by the Rlim (also known as AL022832, AW743871, Ha1r, Rlim1, Rnf12) gene (Gene ID 19820) in murine species. The CLIM/LDB/NLI and RLIM/RNF12 cofactors are transcriptional coregulatory proteins characterized by their ability to bind the LIM domains of nuclear LIM proteins, including LIM homeodomain (LIM-HD) transcription factors. The association of CLIM with LIM-HDs positively regulate their biological and transcriptional activity. On the other hand, the RING finger ubiquitin ligase RLIM acts as a negative regulator of LIM-HD activity by interacting with and targeting CLIM for degradation via the ubiquitin-proteasome pathway.

Specificity

Exhibits no cross-reactivity toward the homologous ubiquitin ligase Rnf6.

Immunogen

Recombinant protein corresponding to mouse RLIM/Rnf12.

Application

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Nuclear Receptors

This Anti-RLIM/Rnf12 Antibody is validated for use in Western Blotting, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP), Immunohistochemistry, Immunofluorescence for the detection of RLIM/Rnf12.

Western Blotting Analysis: A 1:1,000 dilution from a representative lot selectively detected endogenouse RLIM in HEK293 lysate and in vitro transcribed and translated (TNT) RLIM, but not TNT Rnf6.(Courtesy of Dr. Ingolf Bach, University of Massachusetts Medical School).
Western Blotting Analysis: A representative lots detected RLIM in cell and tissue lysates of human, murine, and Chinese hamster species (Jiao, B., et al. (2013). Mol Biol Cell. 24(19):3085-3096; Jiao, B., et al. (2012). Cell. 149(3):630-641; Shin, J., et al. (2010). Nature. 467(7318):977-981; Gungor, C., et al. (2007). Proc. Natl. Acad. Sci. USA. 104(38):15000-15005; Ostendorff, H.P., et al. (2006). Dev Dyn. 235(3):786-791; Kramer, O.H., et al. (2003). EMBO. 22(13):3411-3420; Ostendorff, H.P., et al. (2002). Nature. 416(6876):99-103).
Immunocytochemistry Analysis: A representative lots detected RLIM in cultured murine mammary epithelial cells and MEFs by fluorescent immunocytochemistry (Jiao, B., et al. (2012). Cell. 149(3):630-641; Shin, J., et al. (2010). Nature. 467(7318):977-981).
Immunocytochemistry Analysis: A representative lot detected predominant nuclear expression of RLIM in HeLa, MCF-7, HEK293, and murine pituitary gonadotroph Alpha T3 cells by fluorescent immunocytochemistry (Jiao, B., et al. (2013). Mol Biol Cell. 24(19):3085-3096).
Chromatin Immunoprecipitation Analysis: A representative lots detected RLIM association at the alphaGSU promoter in murine pituitary gonadotroph Alpha T3 cells, as well as estrogen-induced RLIM recruitment to the estrogen response element (ERE) in the promoter region of PS2 and WISP2 in MCF-7 cells (Ostendorff, H.P., et al. (2002). Nature. 416(6876):99-103; Johnsen, S.A., et al. (2009). Cancer Res. 69(1):128-136).
Immunohistochemistry Analysis: A representative lot detected similar stage-dependent expression patterns between RLIM and CLIM1/2 in the developing neural tube during early mouse embryogenesis (Ostendorff, H.P., et al. (2006). Dev Dyn. 235(3):786-791).
Immunohistochemistry Analysis: A representative lot detected a RLIM expression pattern distinct from that of the homologous ubiquitin ligase Rnf6 in E12.5-E13 murine embryonic spinal cord (Tursun, B., et al. (2005). Genes Dev. 19(19):2307-2319).
Immunohistochemistry Analysis: A representative lot detected RLIM-positive cells in breast epidermis and oral cavity epithelium using paraffin-embedded adult human tissue sections (Jiao, B., et al. (2013). Mol Biol Cell. 24(19):3085-3096).
Immunohistochemistry Analysis: A representative lot detected RLIM accumulation during oocyte maturation using paraffin-embedded ovary sections from female mice (Shin, J., et al. (2010). Nature. 467(7318):977-981).
Immunofluorescence Analysis: A representative lot detected RLIM nuclear co-localization with ERalpha in frozen human breast tumor sections by dual fluorescent immunohistochemistry (Johnsen, S.A., et al. (2009). Cancer Res. 69(1):128-136).
Immunoprecipitation Analysis: A representative lot co-immunoprecipitated CLIM and ERalpha with RLIM from MCF-7 lysates (Johnsen, S.A., et al. (2009). Cancer Res. 69(1):128-136).
Immunofluorescence Analysis: A representative lot detected RLIM nuclear co-localization with H3K27me3 immunoreactivity in paraffin-embedded E7-7.5 mouse embryos by dual fluorescent immunohistochemistry (Shin, J., et al. (2014). Nature. 511(7507):86-89).

Quality

Evaluated by Western Blotting in mouse brain tissue lysate.

Western Blotting Analysis: A 1:5,000 of this antibody detected RLIM/Rnf12 in 10 µg of mouse brain tissue lysate.

Target description

~69 kDa observed. Uncharacterized band(s) may appear in some lysates.

Physical form

Rabbit polyclonal serum with 0.05% sodium azide.

Unpurified

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.

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.

biological source: rabbit. Quality Level: 100. antibody form: serum. antibody product type: primary antibodies. clone: polyclonal. species reactivity: hamster, human, mouse. technique(s): ChIP: suitable, immunocytochemistry: suitable, immunofluorescence: suitable, immunohistochemistry: suitable, western blot: suitable. NCBI accession no.: NP_035406. UniProt accession no.: Q9WTV7. shipped in: dry ice. target post-translational modification: unmodified. Gene Information: human ... RLIM(51132). Storage Class Code: 12 - Non Combustible Liquids. WGK: WGK 1. Flash Point(F): Not applicable. Flash Point(C): Not applicable.