UBQ11 | Ubiquitin protein | positive control

86 €

AS10 307S | protein | positive control

PRODUCT INFORMATION IN PDF

product information

background

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

does not apply

antibody format

does not apply

quantity

50 µl

liquid

storage

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

Western blot (WB)

related products

AS08 307 | UBQ11 | Ubiquitin antibodies

additional information

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
recommended dilution		positive control: a 2.5-5 μl load per well is optimal for most chemiluminescent detection systems.
expected \| apparent MW		10-12 kDa
confirmed reactivity		does not apply
predicted reactivity		does not apply
not reactive in		no confirmed exceptions from predicted reactivity currently known
additional information		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
selected references		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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