Part of Thermo Fisher Scientific

UBC Fractions

UBC Fractions Extract for Ctrl

Details
Class: Extracts
Type: Extract
Species Reactivity: Not Applicable (N/A)
Ordering Information
Pierce UBC Fractions
Product Number Pkg. Size Price Purchase
EP-550 400 µg $320.00
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Storage: -80º C, Avoid Freeze/Thaw Cycles
Form: 400 µg in 50 mM HEPES, containing 1 mM DTT; pH 7.6.


Applications Dilution *
Control (Ctrl) Assay Dependent
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Product Specific Information
The enzyme fractions supplied represent the full compliment of purified conjugation enzymes (E1, E2s, and E3s) that are found in mammalian Fraction II. The enzymes have been tested and shown to work with typical [125I]-labeled substrate proteins such as lysozyme and beta-lactoglobulin. These conjugation fractions contain ubiquitin c-terminal hydrolases. The addition of ubiquitin aldehyde is recommended for the inhibition of UCHs and to improve overall conjugate yield. The supplied fractions do not contain 20S or 26S protein degradation activity. If the substrate being conjugated requires E2 or E3 enzymes not found in Fraction II, the reaction can be supplemented with enzymes from Fraction I. Includes complimentary fractions for 2-5 conjugation reactions depending on conditions.

1. Conjugation Fraction A [prot] = 800 ug/ml, 250 µl
2. Conjugation Fraction B, [prot] = 800 ug/ml, 250 µl
General Information
Proteolytic degradation is critical to the maintenance of appropriate levels of short-lived and regulatory proteins as important and diverse as those involved in cellular metabolism, heat shock and stress response, antigen presentation, modulation of cell surface receptors and ion channels, cell cycle regulation, transcription, and signalling factors. The ubiquitin-proteasome pathway deconstructs most proteins in the eukaryotic cell cytosol and nucleus. Others are degraded via the vacuolar pathway which includes endosomes, lysosomes, and the endoplasmic reticulum.

The 26S proteasome is an ATP-dependent, multisubunit (~31), barrel-shaped molecular machine with an apparent molecular weight of ~2.5 MDa. It consists of a 20S proteolytic core complex which is crowned at one or both ends by 19S regulatory subunit complexes. The 19S regulatory subunits recognize ubiquitinated proteins and play an essential role in unfolding and translocating targets into the lumen of the 20S subunit. An enzymatic cascade is responsible for the attachment of multiple ubiquitin molecules to lysine residues of proteins targeted for degradation. Several genetic diseases are associated with defects in the ubiquitin-proteasome pathway. Some examples of affected proteins include those linked to cystic fibrosis, Angelman’s syndrome, and Liddle syndrome.
(This product is for In Vitro experimental use only. Not for resale without express authorization.)