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product information
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| background |
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Ubiquitin is a highly conserved regulatory protein expressed in all eukaryotic tissues. Originally this protein was called: Ubiquitous Immunopoietic Polypeptide. Its function is labeling of proteins for degradation through ubiquitin proteasome system (UPS). Source of positive control, ubiquitin protein: full length recombinant Arabidopsis thaliana ubiquitin, ubq11, with N-terminal 1XHis-3XHA-tag; locus accession: At4g05050 |
| immunogen |
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does not apply |
| antibody format |
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| quantity |
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| storage |
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store protein standard at -20°C; Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tubes. |
| tested applications |
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Western blot (WB) |
| related products |
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AS08 307 | UBQ11 | Ubiquitin antibodies |
| additional information |
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Protein is provided in Laemli SDS loading buffer and should be heated up in 98°C for 5 minutes before loading on the gel. |
application information
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| recommended dilution |
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positive control: a 2.5-5 μl load per well is optimal for most chemiluminescent detection systems. |
| expected | apparent MW |
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10-12 kDa |
| confirmed reactivity |
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does not apply |
| predicted reactivity |
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does not apply |
| not reactive in |
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no confirmed exceptions from predicted reactivity currently known |
| additional information |
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Concentration: 0.8 µg/µl. Recommended load per gel to allow Coomasie staining and western blot detection is 2.5-5 µl. 1x SDS Loading Buffer contains: - 50 mM Tris-HCl pH 6.8
- 2% SDS
- 10% glycerol
- 1% β-mercaptoethanol
- 12.5 mM EDTA
- 0.02% bromophenol blue
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| selected references |
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to be added when available |
application example 5 μg of recombinant protein from Phytophora sp. (1), human (2), Arabidopsis thaliana His-tagged ubiquitin (3), Arabidopsis thaliana His-tagged SUMO protein (4), was separated on 15% PAA gel and blotted on PVDF membrane. Filters were blocked in 5% milk for 1h, incubated with 1: 10 000 anti-ubiquitin antibody (1h), followed by incubation with 1: 15 000 secondary anti-rabbit antibodies(1h) coupled with HRP and visualization (10 seconds exposure) with standard ECL. | | Technical note: It is very difficult to detect ubiquitin monomers in total cell extracts due to a great abundance of poly and multi-ubiquitinated proteins. Recommended is size separation of protein extracts before gel electrophoresis focused on good resolution of region between 6-10 kDa. |
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