2 μg Ubiquitin, AF647-Conjugated run on 4-12% SDS-PAGE gel under reducing conditions, then visualized with Colloidal Coomassie Blue Stain.

Time-dependent assembly of K63-linked polyubiquitin chains using AF647-labeled ubiquitin and the UBE2V1/UBE2N E2 complex. Reactions were resolved by SDS-PAGE and imaged by fluorescence detection on an Invitrogen iBright 1500 Imaging System (ex: 608-632 nm; em: 675-720 nm). Lane 1: no ATP control; lane 2: +ATP, 4 h incubation; lane 3: +ATP, 18 h incubation. Progressive accumulation of higher molecular weight AF647-ubiquitin species in the presence of ATP is consistent with time-dependent formation of K63-linked ubiquitin chains.

Recombinant Human Ubiquitin, AF647-Conjugated

Ub/Ubl's

Product Code: TUB-122

Verified Applications: AF647-conjugated MGCGS-ubiquitin enables fluorescence monitoring of in vitro ubiquitin activation, transfer, and conjugation through E1, E2, and E3 enzyme cascades. Its far-red, photostable fluorophore also supports sensitive deubiquitylase assays for quantitative analysis of ubiquitin pathway dynamics.

Activity: Verified in Ubiquitin Charging Assay.

Purity: 95 %

Concentration: 30 – 50 μM

For Research Use Only (RUO)

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Note: To maintain product quality, larger quantities may be packaged as multiple smaller aliquots. The 1 mg size is generally supplied in a single vial.

UniProt #: Ub, AF647

Predicted Molecular Weight: 9.9 kDa

Tag: Untagged

Background and Additional Info

Background: An AF647-conjugated MGCGS-ubiquitin is a site-specifically labeled ubiquitin derivative engineered to carry a single fluorophore at an N-terminal cysteine residue. The MGCGS extension provides a solvent-accessible thiol for selective conjugation, ensuring homogeneous labeling without perturbing the native ubiquitin fold or its recognition surfaces. The far-red AF647 dye (Ex 650 nm / Em 665 nm) enables high-sensitivity detection with low background and excellent photostability, making this construct well-suited for fluorescence-based assays. It is particularly useful in monitoring ubiquitin conjugation cascades (E1/E2/E3 activity), tracking ubiquitin transfer in real time, and studying deubiquitinase (DUB) activity through loss or redistribution of fluorescence. The defined labeling strategy preserves biochemical functionality while providing a robust optical handle for quantitative and kinetic analyses.

Shipping: The product is shipped with cold packs or dry ice. Upon receipt, store it immediately at the temperature recommended below.

Storage and Stability: Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Aliquot and store ≤ -70 °C (stable for 48 months from date of receipt). This product is light sensitive and should be protected from prolonged light exposure.

Download the Datasheet

Protein Sequence: MG(C*AF647)GSMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG