Anti-Nuclear Spliceosomes Antibody, clone 780-3

Most eukaryotic genes are expressed as pre-mRNAs that are converted to mRNA by splicing. In this process noncoding sequences (introns) are removed, and coding sequences (exons) are ligated together. Some exons are known to be constitutively spliced and are present in every mRNA produced from a given pre-mRNA. However, many are alternatively spliced to generate variable forms of mRNA from a single pre-mRNA species. Nuclear pre-mRNA splicing is catalyzed by the spliceosome that is a multi-megadalton ribonucleoprotein (RNP) complex. It removes introns from nuclear pre-mRNA. The conformation and composition of spliceosome is shown to be dynamic, which is critical for its high degree of accuracy and flexibility. In most eukaryotes two unique spliceosomes have been described: the U2 -dependent spliceosome that catalyzes the removal of U2-type of introns and the U12-dependent spliceosome that is present only in a subset of eukaryotes and splices U12-type of intron. Introns are removed by two consecutive transesterification reactions. In the first reaction the 2 OH group of the branch adenosine of the intron carries out a nucleophilic attack on the 5 ss, which results in cleavage at this site and ligation of the 5 end of the intron to the branch adenosine, forming a lariat structure. In the second reaction, the 3 ss is attacked by the 3 OH group of the 5 exon, which leads to the ligation of the 5 and 3 exons forming the mRNA, and release of the intron. (Ref.: Matera, AG., and Wang, Z. (2014). Nat. Rev. Mol. Cell Biol. 15(2); 108-121; Will, CL., and Luhrmann, R. (2011). Cold Spring Harb. Perspect. Biol. 3(7); a003707; Patel, AA., and Steitz, JA. (2003). Nat. Rev. Mol. Cell Biol. 4(12); 960-970).

Clone 780-3 is a mouse monoclonal antibody that detects human Nuclear spliceosomes.

Nuclear preparation from U266 human myeloma cells.

Quality Control Testing

Evaluated by Flow Cytometry in permeabilized HeLa cells.

Flow Cytometry Analysis: 2 μg of this antibody detected Nuclear Spliceosomes in one million permeabilized HeLa cells.

Tested Applications

Flow Cytometry Analysis: A representative lot detected Nuclear Spliceosomes in Flow Cytometry applications (Clevenger, C.V., et al. (1985). Cytometry. 6(3): 208-14; Bauer, K.D., et al. (1986). Cancer Res. 46(5): 2428-34; Clevenger, C.V., et al. (1987). Cytometry. 8(3): 280-6).

Immunofluorescence Analysis: A representative lot detected Nuclear Spliceosomes in Immunofluorescence applications (Clevenger, C.V., et al. (1984). J Histochem Cytochem. 32(7): 757-65; Clevenger, C.V., et al. (1985). Cytometry. 6(3): 208-14; Bauer, K.D., et al. (1986). Cancer Res. 46(5): 2428-34; Clevenger, C.V., et al. (1987). Cytometry. 8(3):280-6; Clevenger, C.V., et al. (1987). Cytometry. 8(3): 280-6).

Western Blotting Analysis: A representative lot detected Nuclear Spliceosomes in Western Blotting applications (Clevenger, C.V., et al. (1987). Cytometry. 8(3): 280-6; Kumar, P., et al. (2012). PLoS One. 7(8): e42712).

Electron Microscopy: A representative lot detected Nuclear Spliceosomes in Electron Microscopy applications (Clevenger, C.V., et al. (1984). J Histochem Cytochem. 32(7): 757-65; Bauer, K.D., et al. (1986). Cancer Res. 46(5): 2428-34; Clevenger, C.V., et al. (1987). Cytometry. 8(3): 280-6).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-Nuclear Spliceosomes, clone 780-3, Cat. No. MAB1286-I, is a mouse monoclonal antibody that detects Nuclear spliceosomes and is tested for use in Electron Microscopy, Flow Cytometry, Immunofluorescence, and Western Blotting.

Purified mouse monoclonal antibody IgM in PBS without preservatives.

Store at -10°C to -25°C. 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.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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.