Recombinant Anti-RBX1 antibody [EPR6850(B)] (ab133565)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6850(B)] to RBX1
- Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RBX1 antibody [EPR6850(B)]
See all RBX1 primary antibodies -
Description
Rabbit monoclonal [EPR6850(B)] to RBX1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IP: Jurkat whole cell lysate. Flow Cyt: Jurkat cells. ICC/IF: HeLa cells. IHC-P: Rat and mouse breast tissue. Human lung carcinoma tissue. WB: Jurkat whole cell lysate. Mouse and rat heart 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 -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 2.18 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6850(B) -
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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133565 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) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified format use at 1/100 to 1/1000 dilution |
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IP |
1/10 - 1/100.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
For unpurified format use 1/100 to 1/250 dilution |
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WB |
1/5000. Detects a band of approximately 13 kDa (predicted molecular weight: 12 kDa).
For unpurified format use 1/1000 to 1/10000 dilution |
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ICC/IF | (1) |
1/50.
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Notes |
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Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified format use at 1/100 to 1/1000 dilution |
IP
1/10 - 1/100. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. For unpurified format use 1/100 to 1/250 dilution |
WB
1/5000. Detects a band of approximately 13 kDa (predicted molecular weight: 12 kDa). For unpurified format use 1/1000 to 1/10000 dilution |
ICC/IF
1/50. |
Target
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Function
E3 ubiquitin ligase component of multiple cullin-RING-based E3 ubiquitin-protein ligase complexes which mediate the ubiquitination and subsequent proteasomal degradation of target proteins, including proteins involved in cell cycle progression, signal transduction, transcription and transcription-coupled nucleotide excision repair. The functional specificity of the E3 ubiquitin-protein ligase complexes depends on the variable substrate recognition components. As a component of the CSA complex promotes the ubiquitination of ERCC6 resulting in proteasomal degradation. Through the RING-type zinc finger, seems to recruit the E2 ubiquitination enzyme, like CDC34, to the complex and brings it into close proximity to the substrate. Probably also stimulates CDC34 autoubiquitination. May be required for histone H3 and histone H4 ubiquitination in response to ultraviolet and for subsequent DNA repair. Promotes the neddylation of CUL1, CUL2, CUL4 and CUL4 via its interaction with UBE2M. -
Tissue specificity
Widely expressed. -
Pathway
Protein modification; protein ubiquitination. -
Sequence similarities
Belongs to the RING-box family.
Contains 1 RING-type zinc finger. -
Domain
The RING-type zinc finger domain is essential for ubiquitin ligase activity. It coordinates an additional third zinc ion. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 9978 Human
- Entrez Gene: 56438 Mouse
- Entrez Gene: 300084 Rat
- Omim: 603814 Human
- SwissProt: P62877 Human
- SwissProt: P62878 Mouse
- Unigene: 474949 Human
- Unigene: 29405 Mouse
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Alternative names
- BA554C12.1 antibody
- E3 ubiquitin-protein ligase RBX1 antibody
- FLJ60363 antibody
see all
Images
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All lanes : Anti-RBX1 antibody [EPR6850(B)] (ab133565) at 1/5000 dilution
Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat heart lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 12 kDa
Observed band size: 13 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Image produced using the purified version.
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Purified ab133565 at 1:20 dilution (1µg) immunoprecipitating RBX1 in Jurkat whole cell lysate.
Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg.
Lane 2 (+): ab133565 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133565 in Jurkat whole cell lysate.VeriBlot for IP Detection Reagent (HRP)(ab131366) (1:1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 13 kDa -
Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling RBX1 with Purified ab133565 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RBX1 with Purified ab133565 at 1:50 dilution (3.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat breast tissue sections labeling RBX1 with Purified ab133565 at 1:2000 dilution (0.099 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse breast tissue sections labeling RBX1 with Purified ab133565 at 1:2000 dilution (0.099 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling RBX1 with Purified ab133565 at 1:2000 dilution (0.099 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (13)
ab133565 has been referenced in 13 publications.
- Gupta S et al. Functional Characterization of Non-Ankyrin Repeat Domains of Orientia tsutsugamushi Ank Effectors Reveals Their Importance for Molecular Pathogenesis. Infect Immun 90:e0062821 (2022). PubMed: 35435726
- Zhang Q et al. Identification and Validation of Novel Potential Pathogenesis and Biomarkers to Predict the Neurological Outcome after Cardiac Arrest. Brain Sci 12:N/A (2022). PubMed: 35884735
- Wu Q et al. ROC1 promotes the malignant progression of bladder cancer by regulating p-I?Ba/NF-?B signaling. J Exp Clin Cancer Res 40:158 (2021). PubMed: 33962660
- Adcox HE et al. Orientia tsutsugamushi Nucleomodulin Ank13 Exploits the RaDAR Nuclear Import Pathway To Modulate Host Cell Transcription. mBio 12:e0181621 (2021). PubMed: 34340535
- Lanzafame M et al. Cockayne syndrome group A and ferrochelatase finely tune ribosomal gene transcription and its response to UV irradiation. Nucleic Acids Res 49:10911-10930 (2021). PubMed: 34581821