Home Shop UBIQAPTURE-Q® kit

UBIQAPTURE-Q^® kit

BML-UW8995A

Datasheet
Manual
SDS
CofA

Already have an account?

Market-leading tool for isolation and detection of ubiquitinylated proteins.

Efficient tool for the selective isolation of ubiquitinylated proteins
High binding affinity matrix, able to bind both monoubiquitinated and polyubiquitinated proteins
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.

Shipping: Available products typically ship within 24/48h, via priority shipping.

Do you need support? Contact Customer Service or Technical Support.

Online Account
Access or Create Your Account

UBIQAPTURE-Q kit Western Blot — 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.

Product Details

Application Notes	Uses: 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. Capture and analysis of specific ubiquitinylated protein conjugates of interest from particular cell/tissue lysates. 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). 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). 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.
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.
Quantity	Provides sufficient material for 20 x 20μl reactions.

Application Notes

Uses:

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.
Capture and analysis of specific ubiquitinylated protein conjugates of interest from particular cell/tissue lysates.
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).
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).
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.

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.

Quantity

Provides sufficient material for 20 x 20μl reactions.

Handling & Storage

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.
Long Term Storage	-80°C
Shipping	Dry Ice

Regulatory Status	RUO – Research Use Only

Chronic social defeat stress induces the down-regulation of the Nedd4L-GLT-1 ubiquitination pathway in the prefrontal cortex of mice: Kosuge, A., Kunisawa, K., et al.; J. Neurochem. 168, 2479 (2024), Abstract
Susceptibility of Toxoplasma gondii to autophagy in human cells relies on multiple interacting parasite loci: N. Rinkenberger, et al.; Mbio , e0259523 (2023), Abstract
Cotranslational N-degron masking by acetylation promotes proteome stability in plants: E. Linster, et al.; Nat. Commun. 13, 810 (2022), Abstract
Mistargeting of aggregation prone mitochondrial proteins activates a nucleus-mediated posttranscriptional quality control pathway in trypanosomes: Dewar, C. E., Oeljeklaus, S., et al.; Nat. Commun. 13, 3084 (2022), Abstract
SPRTN and TDP1/TDP2 Independently Suppress 5-Aza-2′-deoxycytidine-Induced Genomic Instability in Human TK6 Cell Line: T. Nakano, et al.; Chem. Res. Toxicol. 35, 2059 (2022), Abstract
Parkin-independent mitophagy via Drp1-mediated outer membrane severing and inner membrane ubiquitination: Y. Oshima, et al.; J. Cell Biol. 220, 202006043 (2021), Abstract
Benzo[a]pyrene represses DNA repair through altered E2F1/E2F4 function marking an early event in DNA damage-induced cellular senescence: S. Allmann, et al.; Nucleic Acids Res. 48, 12085 (2020), Abstract — Full Text
Ubiquitination of the Cytoplasmic Domain of Influenza A Virus M2 Protein Is Crucial for Production of Infectious Virus Particles: W.C. Su, et al.; J. Virol. 92, e01972 (2018), Abstract — Full Text
TRIM24 mediates the interaction of the retinoic acid receptor alpha with the proteasome: M. Carrier, et al.; FEBS Lett. 592, 1426 (2018), Abstract — Full Text
Human Cytomegalovirus Tegument Protein pp65 (pUL83) Dampens Type I Interferon Production by Inactivating the DNA Sensor cGAS without Affecting STING: M. Biolatti, et al.; J. Virol. 92, e01774 (2018), Abstract — Full Text