Images
Immunofluorescence

Immunofluorescent analysis of HeLa cells stained with rabbit pAb to ubiquitin, RA22154, dilution 1:1,000 in red, and costained with chicken pAb to vimentin in green. The blue is DAPI staining of nuclear DNA. [A] Control HeLa cells maintained in normal medium, [B] HeLa cells treated with 10µM of the proteasome inhibitor lactacystin (Lc) for 24 hours. Proteasomal inhibition leads to formation of strongly ubiquitin positive cytoplasmic inclusions.

Immunofluorescence

Immunofluorescent analysis of HeLa cells stained with rabbit pAb to ubiquitin, RA22154, dilution 1:1,000 in red, and costained with chicken pAb to vimentin in green. The blue is DAPI staining of nuclear DNA. [A] Control HeLa cells maintained in normal medium, [B] HeLa cells treated with 10µM of the proteasome inhibitor lactacystin (Lc) for 24 hours. Proteasomal inhibition leads to formation of strongly ubiquitin positive cytoplasmic inclusions.

Western Blot

Western blot analysis of HEK293 cell lysates using rabbit pAb to ubiquitin, RA22154, dilution 1:5,000 in green and mouse mAb to β-Tubulin in red, used as a loading control. [1] protein standard (red), [2] cells maintained in normal medium, [3] cells treated with proteasome inhibitor lactacystin (Lc) at 10µM for 16 hours. The smear detected above the 200kDa standard represents accumulations of ubiquitinated proteins in the Lc treated cells.

Western Blot

Western blot analysis of HEK293 cell lysates using rabbit pAb to ubiquitin, RA22154, dilution 1:5,000 in green and mouse mAb to β-Tubulin in red, used as a loading control. [1] protein standard (red), [2] cells maintained in normal medium, [3] cells treated with proteasome inhibitor lactacystin (Lc) at 10µM for 16 hours. The smear detected above the 200kDa standard represents accumulations of ubiquitinated proteins in the Lc treated cells.

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