Recombinant Anti-USP14/TGT antibody [EPR15943] - C-terminal (ab192618)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR15943] to USP14/TGT - C-terminal
- Suitable for: Flow Cyt (Intra), WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-USP14/TGT antibody [EPR15943] - C-terminal
See all USP14/TGT primary antibodies -
Description
Rabbit monoclonal [EPR15943] to USP14/TGT - C-terminal -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HCT-116, HeLa, Jurkat, 293T, C6 and RAW 264.7 cell lysates; HeLa and Jurkat cells. WB: Wild-type HeLa cell lysate, USP14 knockout HeLa cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR15943 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab192618 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/70.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
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ICC/IF |
1/70.
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Notes |
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Flow Cyt (Intra)
1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa). |
ICC/IF
1/70. |
Target
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Function
Proteasome-associated deubiquitinase which releases ubiquitin from the proteasome targeted ubiquitinated proteins. Ensures the regeneration of ubiquitin at the proteasome. Is a reversibly associated subunit of the proteasome and a large fraction of proteasome-free protein exists within the cell. Required for the degradation of the chemokine receptor CXCR4 which is critical for CXCL12-induced cell chemotaxis. Serves also as a physiological inhibitor of endoplasmic reticulum-associated degradation (ERAD) under the non-stressed condition by inhibiting the degradation of unfolded endoplasmic reticulum proteins via interaction with ERN1. Indispensable for synaptic development and function at neuromuscular junctions (NMJs). -
Sequence similarities
Belongs to the peptidase C19 family. USP14/UBP6 subfamily.
Contains 1 ubiquitin-like domain. -
Cellular localization
Cytoplasm. Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 9097 Human
- Entrez Gene: 59025 Mouse
- Entrez Gene: 291796 Rat
- Omim: 607274 Human
- SwissProt: P54578 Human
- SwissProt: Q9JMA1 Mouse
- Unigene: 464416 Human
- Unigene: 707058 Human
see all -
Alternative names
- Deubiquitinating enzyme 14 antibody
- TGT antibody
- tRNA guanine transglycosylase 60 kD subunit antibody
see all
Images
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All lanes : Anti-USP14/TGT antibody [EPR15943] - C-terminal (ab192618) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : USP14 CRISPR-Cas9 edited HeLa cell lysate
Lane 3 : USP14 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 56 kDa
Observed band size: 63 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-USP14/TGT antibody [EPR15943] - C-terminal staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab192618 was shown to bind specifically to USP14/TGT. A band was observed at 63 kDa in wild-type HeLa cell lysates with no signal observed at this size in USP14 CRISPR-Cas9 edited cell line ab266854 (CRISPR-Cas9 edited cell lysate ab257787). The band observed in the CRISPR-Cas9 edited lysate lane below 63 kDa is likely to represent a truncated form of USP14/TGT. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and USP14 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Immunofluorescent analysis of HeLa cells (4% Paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized) labeling USP14/TGT with ab192618 at 1/70 dilution (5 μg/ml). A Goat anti rabbit IgG (Alexa Fluor488) at 1/400 dilution (ab150077) was used as secondary antibody. Counterstain: DAPI.
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Intracellular flow cytometric analysis of Jurkat cells (2% paraformaldehyde-fixed)labeling USP14/TGT with ab192618 at 1/70 dilution (red) or a rabbit IgG (negative) (black), unlabeled cells (blue) followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.
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All lanes : Anti-USP14/TGT antibody [EPR15943] - C-terminal (ab192618) at 1/2000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HCT-116 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 56 kDa -
All lanes : Anti-USP14/TGT antibody [EPR15943] - C-terminal (ab192618) at 1/2000 dilution
Lane 1 : C6 cell lysate
Lane 2 : RAW 264.7 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 56 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (6)
ab192618 has been referenced in 6 publications.
- Guo S et al. TRIB2 modulates proteasome function to reduce ubiquitin stability and protect liver cancer cells against oxidative stress. Cell Death Dis 12:42 (2021). PubMed: 33414446
- Zhang Y et al. Lidocaine inhibits the proliferation and invasion of hepatocellular carcinoma by downregulating USP14 induced PI3K/Akt pathway. Pathol Res Pract N/A:152963 (2020). PubMed: 32471606
- Xu J et al. SPAG5-AS1 inhibited autophagy and aggravated apoptosis of podocytes via SPAG5/AKT/mTOR pathway. Cell Prolif 53:e12738 (2020). PubMed: 31957155
- Xu L et al. IU1 suppresses proliferation of cervical cancer cells through MDM2 degradation. Int J Biol Sci 16:2951-2963 (2020). PubMed: 33061808
- Yu F et al. Tumor suppressive microRNA-124a inhibits stemness and enhances gefitinib sensitivity of non-small cell lung cancer cells by targeting ubiquitin-specific protease 14. Cancer Lett 427:74-84 (2018). WB . PubMed: 29702194
- Fletcher AJ et al. Trivalent RING Assembly on Retroviral Capsids Activates TRIM5 Ubiquitination and Innate Immune Signaling. Cell Host Microbe 24:761-775.e6 (2018). PubMed: 30503508