Product nameAnti-RNF144A antibody
See all RNF144A primary antibodies
DescriptionRabbit polyclonal to RNF144A
Tested applicationsSuitable for: IHC-P, ICC/IF, WB, ELISAmore details
Species reactivityReacts with: Human, Rhesus monkey
Predicted to work with: Mouse
Synthetic peptide corresponding to Human RNF144A (internal sequence).
- extracts from COLO cells This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HeLa. IHC-P: Human prostate FFPE tissue sections.
Previously labelled as RNF144.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 50% Glycerol, 0.87% Sodium chloride, PBS
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab75054 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||1/500 - 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 33 kDa).|
FunctionE3 ubiquitin-protein ligase which accepts ubiquitin from E2 ubiquitin-conjugating enzymes UBE2L3 and UBE2L6 in the form of a thioester and then directly transfers the ubiquitin to targeted substrates.
PathwayProtein modification; protein ubiquitination.
Sequence similaritiesBelongs to the RBR family. RNF144 subfamily.
Contains 1 IBR-type zinc finger.
Contains 2 RING-type zinc fingers.
- Information by UniProt
- KIAA0161 antibody
- Probable E3 ubiquitin protein ligase RNF144A antibody
- Probable E3 ubiquitin-protein ligase RNF144A antibody
All lanes : Anti-RNF144A antibody (ab75054) at 1/500 dilution
Lane 1 : extracts from COLO cells
Lane 2 : extracts from COLO cells with immunising peptide at 5 µg
Lysates/proteins at 5 µg per lane.
Predicted band size: 33 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?
Additional bands at: 86 kDa. We are unsure as to the identity of these extra bands.
IHC image of RNF144A staining in human prostate formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75054, 1µg/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.
ICC/IF image of ab75054 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 ab75054 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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 product has been referenced in:
- Chen X et al. DHHC protein family targets different subsets of glioma stem cells in specific niches. J Exp Clin Cancer Res 38:25 (2019). Read more (PubMed: 30658672) »
- Ho SR et al. Regulation of RNF144A E3 Ubiquitin Ligase Activity by Self-association through Its Transmembrane Domain. J Biol Chem 290:23026-38 (2015). Read more (PubMed: 26216882) »