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Recombinant full length protein corresponding to Human Ubiquitin.
This antibody stains the periphery of senile plaques and of neurofibrillary tangles in Alzheimer's disease brain, the Lewy bodies in Parkinson's disease brain, and the Mallory bodies in alcoholic liver disease.
Our Abpromise guarantee covers the use of ab7780 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.
1/500 - 1/1000.
|IHC-FoFr||1/50 - 1/100.|
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Boil tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 8.5 kDa. Used at a dilution of 1/2000 B16F10 murine melanoma cell line (see Abreview). Predicted molecular weight: 8.5 kDa.|
|IP||Use at an assay dependent concentration.|
Image courtesy of an anonymous Abreview.
ab7780 used in Western Blot. Whole cell lysate prepared from human lung cancer cell lines was loaded at 100000 cells. The gel was run under reduced, denaturing conditions. 5% milk used as the blocking agent. ab7780 used at a 1/1000 dilution, incubated for 16 hours at 4°C in 5% BSA in TBS-Tween (0.1%). The secondary was an HRP conjugated mouse anti-rabbit, used at a 1/10,000 dilution.Specific bands seen : 16, 24, 32, 50, 60, kDa.
This image is courtesy of an Abreview submitted by Armen PetrosyanUbiquitin western blot of the complexes pulled down with anti-c-Myc antibodies from the lysates of cells treated with DMSO or MG-132. Treatment with the proteasome inhibitor, MG-132, increases ubiquitinated protein; seen as several bands between 55-72 kDa.
ab7780 staining Ubiquitin in Panc-1 cells by Immunocytochemistry/ Immunofluorescence.Cells were fixed in formaldehyde, blocked with 1% serum for 1 hour at 22°C and then incubated with ab7780 at a 1/2000 dilution for 1 hour at 22°C. The secondary was used at a 1/200 dilution.Green - Golgi residental proteins C2GnT-M.Red -Ubiquitin.Blue - nucleus staining by DAPI.
ab7780 staining ubiquitin in cells transfected with FGAMS-EGFP by ICC/IF (immunocytochemistry/immunofluorescence). Cells were fixed with 3.7% methanol-free formaldehyde at 37°C for 15-20 minutes, blocked with 5% goat serum in PBS-T buffer for 30-60 minutes at room temperature. An Alexa Fluor® 594-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody
ICC/IF image of ab7780 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7780, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse IgG (H+L) ab150113) used at a 1/1000 dilution for 1h. Alexa Fluor© 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43æM.
ab7780 (1/100) staining Ubiquitin in paraffin-embedded brain sections of a patient with Alzheimer's disease.
ab7780 staining ubiquitin in Mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/90) for 16 hours at 4°C. A HRP-conjugated Goat anti-rabbit polyclonal (1/1000) was used as the secondary antibody.
ab7780 at a dilution of 1/1000, staining Ubiquitin in neurons (Alexa 488 secondary at 1/2000) on 30µm coronal rat brain (ab29475) tissue sections in free floating IHC (see protocol link for detailed description). Image shows neuronal staining observed with [A] 20x objective and [B] 40x objective.
NB: No labeling observed following omission of primary antibody.
Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).
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