Key features and details
- Rabbit polyclonal to HUWE1/Mule
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-HUWE1/Mule antibody
See all HUWE1/Mule primary antibodies
DescriptionRabbit polyclonal to HUWE1/Mule
Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Chimpanzee, Gorilla, Orangutan
Synthetic peptide corresponding to Human HUWE1/Mule aa 2250-2300.
Database link: Q7Z6Z7
This product was previously labelled as HUWE1
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.09% Sodium azide
Constituents: 1.815% Tris, 1.764% Sodium citrate, 0.021% PBS
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab70161 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||1/1000 - 1/5000. Detects a band of approximately >460 kDa (predicted molecular weight: 481 kDa).|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
FunctionE3 ubiquitin-protein ligase which mediates ubiquitination and subsequent proteasomal degradation of target proteins. Regulates apoptosis by catalyzing the polyubiquitination and degradation of MCL1. Mediates monoubiquitination of DNA polymerase beta (POLB) at 'Lys-41', 'Lys-61' and 'Lys-81', thereby playing a role in base-excision repair. Also ubiquitinates the p53/TP53 tumor suppressor and core histones including H1, H2A, H2B, H3 and H4. Binds to an upstream initiator-like sequence in the preprodynorphin gene. Regulates neural differentiation and proliferation by catalyzing the polyubiquitination and degradation of MYCN. May regulate abundance of CDC6 after DNA damage by polyubiquitinating and targeting CDC6 to degradation.
Tissue specificityWeakly expressed in heart, brain and placenta but not in other tissues. Expressed in a number of cell lines, predominantly in those from colorectal carcinomas.
PathwayProtein modification; protein ubiquitination.
Involvement in diseaseDefects in HUWE1 are the cause of mental retardation syndromic X-linked Turner type (MRXST) [MIM:300706]; also known as mental retardation and macrocephaly syndrome. MRXST shows clinical variability. Associated phenotypes include macrocephaly and variable contractures.
A chromosomal microduplication involving HUWE1 and HSD17B10 is the cause of mental retardation X-linked type 17 (MRX17) [MIM:300705]; also known as mental retardation X-linked type 31 (MRX31). Mental retardation is characterized by significantly sub-average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. In contrast to syndromic or specific X-linked mental retardation which also present with associated physical, neurological and/or psychiatric manifestations, intellectual deficiency is the only primary symptom of non-syndromic X-linked mental retardation.
Sequence similaritiesBelongs to the TOM1/PTR1 family.
Contains 1 HECT (E6AP-type E3 ubiquitin-protein ligase) domain.
Contains 1 UBA domain.
Contains 1 UIM (ubiquitin-interacting motif) repeat.
Contains 1 WWE domain.
DomainThe HECT domain mediates inhibition of the transcriptional activity of p53.
modificationsPhosphorylated on tyrosine; phosphorylation is probably required for its ability to inhibit TP53 transactivation.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationCytoplasm. Nucleus. Mainly expressed in the cytoplasm of most tissues, except in the nucleus of spermatogonia, primary spermatocytes and neuronal cells (By similarity). Predominantly cytosolic or perinuclear in some colorectal carcinoma cells.
- Information by UniProt
- ARF binding protein 1 antibody
- ARF BP1 antibody
- ARF-binding protein 1 antibody
All lanes : Anti-HUWE1/Mule antibody (ab70161) at 1 µg/ml
Lane 1 : 293T whole cell lysate at 50 µg
Lane 2 : 293T whole cell lysate at 15 µg
Lane 3 : 293T whole cell lysate at 5 µg
Predicted band size: 481 kDa
Observed band size: >460 kDa why is the actual band size different from the predicted?
Additional bands at: 200 kDa, 300 kDa. We are unsure as to the identity of these extra bands.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling HUWE1/Mule with ab70161 at 1/1000 (1µg/ml). Detection: DAB.
ICC/IF image of ab70161 stained HepG2 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 (ab70161, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.
IHC image of ab70161 staining in human normal heart formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab70161, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab70161 has been referenced in 6 publications.
- Flack JE et al. Wnt-Dependent Inactivation of the Groucho/TLE Co-repressor by the HECT E3 Ubiquitin Ligase Hyd/UBR5. Mol Cell 67:181-193.e5 (2017). PubMed: 28689657
- Byrne R et al. Activity-Based Probes for HECT E3 Ubiquitin Ligases. Chembiochem 18:1415-1427 (2017). PubMed: 28425671
- Park SH et al. Metformin enhances TRAIL-induced apoptosis by Mcl-1 degradation via Mule in colorectal cancer cells. Oncotarget 7:59503-59518 (2016). PubMed: 27517746
- Horikawa I et al. Autophagic degradation of the inhibitory p53 isoform ?133p53a as a regulatory mechanism for p53-mediated senescence. Nat Commun 5:4706 (2014). PubMed: 25144556
- Izumi H & Kaneko Y Trim32 facilitates degradation of MYCN on spindle poles and induces asymmetric cell division in human neuroblastoma cells. Cancer Res 74:5620-30 (2014). PubMed: 25100564
- Chan CH et al. Deciphering the transcriptional complex critical for RhoA gene expression and cancer metastasis. Nat Cell Biol 12:457-67 (2010). WB ; Human . PubMed: 20383141