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Efficient tool for the selective isolation of ubiquitinylated proteins
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High binding affinity matrix, able to bind both monoubiquitinated and polyubiquitinated proteins
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Highly compatible with a wide range of lysate buffer, different sample types (cells, tissues) and a broad range of species
For isolation and enrichment of ubiquitinylated proteins.The UBI-QAPTURE-Q® kit is an efficient tool for the selective isolation of ubiquitinylated proteins. The Kit facilitates the isolation of both mono- and poly-ubiquitinylated proteins (independent of lysine residue chain linkage) from cell extracts, tissue lysates and in vitro assay solutions through use of a high-binding affinity matrix. Captured proteins can be analyzed by Western blotting using the highly sensitive ubiquitin-conjugate specific antibody provided, using antibodies to specific proteins of interest, or eluted from the matrix for subsequent biochemical characterization. The UBI-QAPTURE-Q® matrix supplied with the kit has superior binding characteristics compared to other commercially available matrices and is compatible with a wide range of lysate buffers and cell/tissue samples from a variety of species.
Western Blot analysis of ubiquitin enrichment of lysate derived ubiquitinylated proteins. Ubiquitin enrichment experiment set-up and run as described in UBIQAPTURE-Q® kit (Prod. No. BML-UW8995A). Ubiquitin-protein conjugates present in starting material, unbound fraction and elution fraction were detected by western blotting using the provided ubiquitin-conjugate specific HRP-linked antibody (Prod. No. ENZ-ABS840HRP) at a dilution of 1:1000. Key: SM = Starting Material, UF = Unbound Fraction, WA = Wash Fraction, and EL = Elution Fraction.
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Product Specification
| Application Notes: | Uses:
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Isolation and detection of the full range of ubiquitinylated protein conjugates (mono-/multi-/poly-ubiquitin modified, lysine linkage independent) from specific cell/tissue lysates of interest, especially low abundance (regulatory) proteins involved in the ubiquitin-proteasome pathway.
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Capture and analysis of specific ubiquitinylated protein conjugates of interest from particular cell/tissue lysates.
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Purification/pull down of ubiquitinylated proteins from cell free in vitro assays; for example to isolate Ub-p53 species from ubiquitinylation assays mediated by HeLa S100 lysate (Prod. No. BML-SW8750).
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Release of free proteins in their active/native form by cleavage of ubiquitin/ubiquitin chains from the UBI-QAPTURE-Q® matrix using deubiquitinylating enzymes such as USP2 (Prod. No. BML-UW9850).
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Release of ubiquitinylated proteins in their active/native form by elution from the UBI-QAPTURE-Q® matrix using, for example, high salt buffer.
Note: Protocol provided covers applications 1-3. Assay set-up can be readily modified for alternative applications by inclusion, omission or substitution of specific components. Kit is not suitable for use with purified proteins. |
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| Quantity: | Provides sufficient material for 20 x 20μl reactions. |
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| Use/Stability: | Upon receipt, the control lysate (Prod. No. BML-UW0130) should be stored at -80°C. All other components should be stored at 4°C. |
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| Shipping: | Shipped on Dry Ice |
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| Long Term Storage: | -80°C |
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| Contents: | UBI-QAPTURE-Q® matrix (Prod. No. BML-UW0125) 0.5 ml settled resin provided; 50% suspension.
Control ubiquitinylated-protein lysate (Ub-lysate) (Prod. No. BML-UW0130) 500 µg (100µl), 5mg/ml in 50mM HEPES, pH 7.6, 1mM DTT.
Mono- and polyubiquitinylated conjugates monoclonal antibody (UBCJ2) (HRP conjugate) (Ubiquitin-conjugates specific HRP-linked antibody solution) (Prod. No. ENZ-ABS840HRP) 25µl. |
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| Regulatory Status: | RUO - Research Use Only |
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