Anti-RNF31/HOIP antibody (ab46322)
Key features and details
- Rabbit polyclonal to RNF31/HOIP
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-RNF31/HOIP antibody
See all RNF31/HOIP primary antibodies -
Description
Rabbit polyclonal to RNF31/HOIP -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse
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Immunogen
Synthetic peptide corresponding to Human RNF31/HOIP aa 1050 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab46321)
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab46322 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 ICC/IF WB Application notesChIP: Use at an assay dependent dilution (PMID 19237537.
ICC/IF: Use at a concentration of 1 µg/ml.
WB: 1/250. Detects a band of approximately 116 kDa (predicted molecular weight: 119 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.Target
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Function
E3 ubiquitin-protein ligase component of the LUBAC complex which conjugates linear polyubiquitin chains in a head-to-tail manner to substrates and plays a key role in NF-kappa-B activation and regulation of inflammation. LUBAC conjugates linear polyubiquitin to IKBKG and RIPK1 and is involved in activation of the canonical NF-kappa-B and the JNK signaling pathways. Linear ubiquitination mediated by the LUBAC complex interferes with TNF-induced cell death and thereby prevents inflammation. LUBAC is proposed to be recruited to the TNF-R1 signaling complex (TNF-RSC) following polyubiquitination of TNF-RSC components by BIRC2 and/or BIRC3 and to conjugate linear polyubiquitin to IKBKG and possibly other components contributing to the stability of the complex. Binds polyubiquitin of different linkage types. -
Tissue specificity
Expressed in both normal and transformed breast epithelial cell lines. -
Sequence similarities
Contains 2 IBR-type zinc fingers.
Contains 3 RanBP2-type zinc fingers.
Contains 1 RING-type zinc finger.
Contains 1 UBA domain. -
Domain
The RanBP2-type zinc fingers mediate the specific interaction with ubiquitin. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 55072 Human
- Entrez Gene: 268749 Mouse
- Omim: 612487 Human
- SwissProt: Q96EP0 Human
- SwissProt: Q924T7 Mouse
- Unigene: 375217 Human
- Unigene: 434081 Human
- Unigene: 23748 Mouse
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Alternative names
- E3 ubiquitin protein ligase RNF31 antibody
- FLJ10111 antibody
- FLJ23501 antibody
see all
Images
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Western blot - Anti-RNF31/HOIP antibody (ab46322)Anti-RNF31/HOIP antibody (ab46322) at 1/250 dilution + Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 119 kDa
Observed band size: 116 kDa why is the actual band size different from the predicted?
Additional bands at: 105 kDa (possible isoform), 50 kDa (possible isoform), 60 kDa. We are unsure as to the identity of these extra bands. -
Immunocytochemistry/ Immunofluorescence - Anti-RNF31/HOIP antibody (ab46322)ICC/IF image of ab46322 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab46322, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) Hek293 cells at 5µg/ml. -
Western blot - Anti-RNF31/HOIP antibody (ab46322)This image is courtesy of an anonymous AbreviewWestern blot analysis of A549 cell lysate (100μg/lane) RNF31/HOIP with ab46322 at 1/1000. A HRP-conjugated Goat anti-rabbit polyclonal (1/10000) was used as the secondary antibody.
lane 1-2 : siCT A549
lane 3-4 : siHOIP A549
Protocols
Datasheets and documents
References (15)
ab46322 has been referenced in 15 publications.
- Oda H et al. Second Case of HOIP Deficiency Expands Clinical Features and Defines Inflammatory Transcriptome Regulated by LUBAC. Front Immunol 10:479 (2019). PubMed: 30936877
- Priem D et al. A20 protects cells from TNF-induced apoptosis through linear ubiquitin-dependent and -independent mechanisms. Cell Death Dis 10:692 (2019). PubMed: 31534131
- Liu Y et al. Down-regulation of MicroRNA-133 predicts poor overall survival and regulates the growth and invasive abilities in glioma. Artif Cells Nanomed Biotechnol 46:206-210 (2018). PubMed: 28376685
- Dziedzic SA et al. ABIN-1 regulates RIPK1 activation by linking Met1 ubiquitylation with Lys63 deubiquitylation in TNF-RSC. Nat Cell Biol 20:58-68 (2018). PubMed: 29203883
- Qiu P et al. MicroRNA-378 regulates cell proliferation and migration by repressing RNF31 in pituitary adenoma. Oncol Lett 15:789-794 (2018). PubMed: 29399147
Images
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Western blot - Anti-RNF31/HOIP antibody (ab46322)Anti-RNF31/HOIP antibody (ab46322) at 1/250 dilution + Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 119 kDa
Observed band size: 116 kDa why is the actual band size different from the predicted?
Additional bands at: 105 kDa (possible isoform), 50 kDa (possible isoform), 60 kDa. We are unsure as to the identity of these extra bands.
-
Immunocytochemistry/ Immunofluorescence - Anti-RNF31/HOIP antibody (ab46322)ICC/IF image of ab46322 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab46322, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) Hek293 cells at 5µg/ml.
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Western blot - Anti-RNF31/HOIP antibody (ab46322) This image is courtesy of an anonymous Abreview
Western blot analysis of A549 cell lysate (100μg/lane) RNF31/HOIP with ab46322 at 1/1000. A HRP-conjugated Goat anti-rabbit polyclonal (1/10000) was used as the secondary antibody.
lane 1-2 : siCT A549
lane 3-4 : siHOIP A549
