Product DescriptiongoogleRabbit anti-Ubiquitin Polyclonal Antibody (Unconjugated), suitable for WB, ICC.
Alternative NamesRPS27A; UBA52; UBB; UBC; Polyubiquitin-B; Polyubiquitin-C;
Application(s)ICC, WB
Antibody HostRabbit
Antibody TypePolyclonal
SpecificityThe specificity of this antibody has been confirmed by WB. This antibody detects ~8.5 kDa Ubiquitin. Human. Predicted to react with other mammalian tissues.
Product DescriptionRabbit anti-Ubiquitin Polyclonal Antibody (Unconjugated), suitable for WB, ICC.
Application(s)ICC, WB
Application DetailsWestern Blotting (WB) and Immunocytochemistry (ICC). A dilution of 1:5,000 - 1:10,000 is recommended for WB. A dilution of 1:500-1,000 is recommended for ICC. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetUbiquitin
SpecificityThe specificity of this antibody has been confirmed by WB. This antibody detects ~8.5 kDa Ubiquitin. Human. Predicted to react with other mammalian tissues.
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 50 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsAfter reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesRPS27A; UBA52; UBB; UBC; Polyubiquitin-B; Polyubiquitin-C;
Scientific BackgroundUbiquitin is a highly conserved 76 amino acid protein with an estimated molecular weight of 8.56 kDa which has a central role in regulated protein degradation. It is a protein modifier which can be covalently attached to target lysines either as a monomer or as a lysine-linked polymer. Several types of polymeric chains can be formed depending on the lysine used for the assembly. Attachment to proteins as a polymer leads to their degradation by the 26S proteosome; a complex, multicatalytic cytosolic and nuclear protease. Attachment to proteins as a monomer or as an alternatively linked polymer does not lead to proteasomal degradation and may be required for numerous functions, including maintenance of chromatic structure, regulation of gene expression, stress response, ribosome biogenesis and DNA repair. Ubiquitin is synthesized as a polyubiquitin precursor with exact head to tail repeats, the number of repeats of which differ between species and strains. In some species there is a final amino-acid after the last repeat, here in bovine a Cys. Some ubiquitin genes contain a single copy of ubiquitin fused to a ribosomal protein (either L40 or S27a).
Left: Immunofluorescent analysis of HeLa cells stained with rabbit antibody to ubiquitin (red, 1:1,000) by Immunocytochemistry. Cells were co-stained with chicken antibody to vimentin (C-1409-50, green, 1:10,000). Blue: DAPI nuclear stain. [A] Control HeLa cells maintained in normal medium; [B] HeLa cells treated with 10 uM of the proteasome inhibitor lactacystin (Lc) for 24 hours. Proteasomal inhibition leads to formation of strongly ubiquitin positive cytoplasmic inclusions [B]. Note the diffuse cytoplasmic ubiquitin staining in control cells [A] and well-defined ubiquitin positive inclusions in the Lc treated cells [B]. Right: Western blot analysis of HEK293 cell lysates using rabbit antibody to ubiquitin (green, 1:5,000). [1] protein standard, [2] cells maintained in normal medium, [3] cells treated with proteasome inhibitor lactacystin (Lc) at 10 um for 16 hours. The blot was simultaneously probed with a mouse antibody to beta-tubulin as loading control (red, lanes 2 and 3). The smear detected above the 200 kDa standard represents accumulations of ubiquitinated proteins in the Lc-treated cells. The prominent band at ~8 kDa corresponds to monoubiquitin.
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