Recombinant Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
![Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-254330-anti-stub1-chip-antibody-epr4447-western-blot.jpg "Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4447] to STUB1/CHIP
- Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-STUB1/CHIP antibody [EPR4447]
See all STUB1/CHIP primary antibodies -
Description
Rabbit monoclonal [EPR4447] to STUB1/CHIP -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human STUB1/CHIP aa 1-100 (N terminal). The exact sequence is proprietary.
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Positive control
- HeLa, MCF-7, 293T and ECV-304 cell lysates; Human skeletal muscle tissue.
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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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4447 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab134064 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 |
|---|---|---|
| WB | (5) |
1/10000 - 1/50000. Predicted molecular weight: 35 kDa.
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| IP | (2) |
1/120.
For unpurified, use 1/10 - 1/100. |
| Flow Cyt (Intra) |
Use at an assay dependent concentration.
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| ICC/IF | (2) |
1/250 - 1/500.
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| IHC-P |
1/100.
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| Notes |
|---|
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WB
1/10000 - 1/50000. Predicted molecular weight: 35 kDa. |
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IP
1/120. For unpurified, use 1/10 - 1/100. |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
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ICC/IF
1/250 - 1/500. |
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IHC-P
1/100. |
Target
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Function
E3 ubiquitin-protein ligase which targets misfolded chaperone substrates towards proteasomal degradation. Ubiquitinates NOS1 in concert with Hsp70 and Hsp40. Modulates the activity of several chaperone complexes, including Hsp70, Hsc70 and Hsp90. Mediates transfer of non-canonical short ubiquitin chains to HSPA8 that have no effect on HSPA8 degradation. Mediates polyubiquitination of DNA polymerase beta (POLB) at 'Lys-41', 'Lys-61' and 'Lys-81', thereby playing a role in base-excision repair: catalyzes polyubiquitination by amplifying the HUWE1/ARF-BP1-dependent monoubiquitination and leading to POLB-degradation by the proteasome. Mediates polyubiquitination of CYP3A4. -
Tissue specificity
Highly expressed in skeletal muscle, heart, pancreas, brain and placenta. Detected in kidney, liver and lung. -
Pathway
Protein modification; protein ubiquitination. -
Sequence similarities
Contains 3 TPR repeats.
Contains 1 U-box domain. -
Domain
The TPR domain is essential for ubiquitination mediated by UBE2D1. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Auto-ubiquitinated; mediated by UBE2D1 and UBE2D2. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 10273 Human
- Entrez Gene: 56424 Mouse
- Entrez Gene: 287155 Rat
- Omim: 607207 Human
- SwissProt: Q9UNE7 Human
- SwissProt: Q9WUD1 Mouse
- Unigene: 592081 Human
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Alternative names
- Antigen NY CO 7 antibody
- Antigen NY-CO-7 antibody
- C terminus of Hsp70-interacting protein antibody
see all
Images
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Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)All lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/20000 dilution (purified)
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
Lane 3 : HEK293 (Human epithelial cell line from embryonic kidney) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 35 kDa
Observed band size: 35 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Immunocytochemistry/ Immunofluorescence - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-253352-anti-stub1-chip-antibody-epr4447-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)Immunofluorescence staining of SH-SY5Y cells with purified ab134064 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab134064 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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![Immunoprecipitation - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-253363-anti-stub1-chip-antibody-epr4447-immunoprecipitation.jpg)
Immunoprecipitation - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)ab134064 (purified) at 1/120 immunoprecipitating STUB1/CHIP in 10 μg HeLa (Lanes 1 and 2, observed at 35 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-148187-anti-stub1-chip-antibody-epr4447-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling STUB1/CHIP with unpurified ab134064 at 1/100 dilution.
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![Flow Cytometry (Intracellular) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-272317-anti-stub1-chip-antibody-epr4447-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)Intracellular Flow Cytometry analysis of SH-SY5Y (Human epithelial cell line from embryonic kidney) cells labeling STUB1/CHIP with purified ab134064 at 1/120 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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![Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-254333-anti-stub1-chip-antibody-epr4447-western-blot.jpg)
Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/10000 dilution (purified) + Mouse brain at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 35 kDa
Observed band size: 35 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-226944-anti-stub1-chip-antibody-epr4447-western-blot.jpg)
Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)This image is courtesy of an anonymous AbreviewAll lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/1000 dilution (unpurified)
Lane 1 : Wild type primary Mouse neurons whole cell lysate
Lane 2 : STUB1 knockout primary Mouse neurons whole cell lysate
Lane 3 : Wild type MEF (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 4 : STUB1 knockout MEF (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 35 kDa
Exposure time: 25 seconds
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![Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-254334-anti-stub1-chip-antibody-epr4447-western-blot.jpg)
Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/50000 dilution (purified) + Rat brain at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 35 kDa
Observed band size: 35 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-148188-anti-stub1-chip-antibody-epr4447-western-blot.jpg)
Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)All lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/10000 dilution (unpurified)
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : MCF-7 (Human breast adenocarcinoma cell line) cell lysate
Lane 3 : 293T cell lysate
Lane 4 : ECV-304 (Human urinary bladder cancer cell line) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 35 kDaECV-304 (derived from T24, discontinued by ATCC)
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![Anti-STUB1/CHIP antibody [EPR4447] (ab134064)](https://www.abcam.com/ps/products/134/ab134064/Images/ab134064-10-benefits-of-recombinant-antibodies.png)
Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
Protocols
Datasheets and documents
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Datasheet download
References (9)
ab134064 has been referenced in 9 publications.
- Liu Y et al. CHIP-mediated CIB1 ubiquitination regulated epithelial-mesenchymal transition and tumor metastasis in lung adenocarcinoma. Cell Death Differ 28:1026-1040 (2021). PubMed: 33082516
- Wu HH et al. Hsp70 acts as a fine-switch that controls E3 ligase CHIP-mediated TAp63 and ?Np63 ubiquitination and degradation. Nucleic Acids Res 49:2740-2758 (2021). PubMed: 33619536
- Chen BH et al. Hsc70/Stub1 promotes the removal of individual oxidatively stressed peroxisomes. Nat Commun 11:5267 (2020). PubMed: 33077711
- Schuster S et al. CHIP mutations affect the heat shock response differently in human fibroblasts and iPSC-derived neurons. Dis Model Mech 13:N/A (2020). PubMed: 33097556
- Zhang L et al. Cimetidine promotes STUB1-mediated degradation of tumoral FOXP3 by activating PI3K-Akt pathway in gastric cancer. Ann Transl Med 8:1304 (2020). PubMed: 33209884
- Dema A et al. Cyclin-Dependent Kinase 18 Controls Trafficking of Aquaporin-2 and Its Abundance through Ubiquitin Ligase STUB1, Which Functions as an AKAP. Cells 9:N/A (2020). PubMed: 32164329
- Wang Y et al. STUB1 is targeted by the SUMO-interacting motif of EBNA1 to maintain Epstein-Barr Virus latency. PLoS Pathog 16:e1008447 (2020). PubMed: 32176739
- Pan C et al. Hsp90B enhances MAST1-mediated cisplatin resistance by protecting MAST1 from proteosomal degradation. J Clin Invest 129:4110-4123 (2019). PubMed: 31449053
- Li H et al. WWP2 is a physiological ubiquitin ligase for phosphatase and tensin homolog (PTEN) in mice. J Biol Chem 293:8886-8899 (2018). PubMed: 29685889
