The differentiation and activation of monocytes (MO) and monocytic cells is modulated by 1alpha,25-dihydroxyvitamin D3 (Vitamin D3). Recently, using mRNA Differential Display technology, Fritsche J, et al have identified a gene which is induced in freshly isolated human blood MO that are cultured for 4 hours with Vitamin D3 (1). This cDNA encodes a putative 16.5kD protein. The product of this gene has structural homology with yeast MMS2p and human CROC-1), which defines a structurally and functionally conserved Ubc-like protein family (2). This protein was called Ubiquitin E2 variant (UBE2V2) or MMS2 (methyl methanesulfonate sensitive 2). The yeast UEV protein, Mms2, is required together with Ubc13 for RAD6/RAD18-dependent postreplicative DNA repair in yeast (3). These proteins also play a central role in the assembly of K63-linked polyubiquitin chains (4,5) UEV/Ubc complexes have been implicated in the assembly of Lys63-linked polyubiquitin chains that act as a novel signal in postreplicative DNA repair and IkBa kinase activation. Recent crystal structure analysis provides direct evidence that the Mms2/Ubc13 heterodimer is necessary for DNA repair (6,7).
Applications:
Suitable for use in Western Blot. Other applications not tested.
Recommended Dilution:
Optimal dilutions to be determined by the researcher.
Positive Control: HL-60
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol (40-50%), aliquot and store at -20 degrees C. Aliquots are stable for at least 3 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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