2 μg Ubiquitin, AF488-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 AF488-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: 455-485 nm; em: 508-557 nm). Lane 1: no ATP control; lane 2: +ATP, 4 h incubation; lane 3: +ATP, 18 h incubation. Progressive accumulation of higher molecular weight AF488-ubiquitin species in the presence of ATP is consistent with time-dependent formation of K63-linked ubiquitin chains.

Recombinant Human Ubiquitin, AF488-Conjugated

Ub/Ubl's

Product Code: TUB-120

Verified Applications: AF488-conjugated MGCGS-ubiquitin enables fluorescence monitoring of in vitro ubiquitin activation, transfer, and conjugation through E1, E2, and E3 enzyme cascades. This bright, 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, AF488

Predicted Molecular Weight: 9.9 kDa

Tag: Untagged

Background and Additional Info

Background: An AF488-conjugated MGCGS-ubiquitin is a site-specifically labeled ubiquitin derivative engineered with an N-terminal cysteine for controlled fluorophore attachment. The MGCGS extension enables selective thiol conjugation, producing a homogeneous, single-label species while preserving the native ubiquitin structure and functional interfaces. The bright green-emitting AF488 dye (Ex 495 nm / Em 519 nm) provides high quantum yield and strong signal intensity, making this construct well-suited for fluorescence microscopy, flow-based assays, and in vitro enzymatic studies. It supports sensitive detection of ubiquitylation cascades, real-time monitoring of ubiquitin transfer, and analysis of deubiquitylase activity, while maintaining compatibility with standard GFP/FITC filter sets.

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.

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Protein Sequence: MG(C*AF488)GSMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG