Product nameAnti-RAI1 antibody
See all RAI1 primary antibodies
DescriptionRabbit polyclonal to RAI1
Tested applicationsSuitable for: ICC/IF, IHC-P, WB, IPmore details
Species reactivityReacts with: Human
Predicted to work with: Chimpanzee, Gorilla, Orangutan
Synthetic peptide corresponding to a region between residue 25 and 75 of Human RAI1 (NP_109590.3).
- Whole cell lysate from HeLa cells. This antibody gave a positive result when used in the following methanol fixed cell lines: HeLa
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.09% Sodium Azide
Constituents: Tris citrate/phosphate buffer, pH 7-8
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab86599 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 5 µg/ml.|
|IHC-P||1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||1/2000 - 1/10000. Predicted molecular weight: 203 kDa.|
|IP||Use at 10 µg/mg of lysate.|
RelevanceRAI1 (retinoid-acid induced protein 1) may be involved in neuronal differentiation. RAI1 is highly similar to its mouse counterpart and is expressed at high levels mainly in neuronal tissues. RAI1 has a polymorphic polyglutamine tract in it's N-terminal domain. Expression of the mouse counterpart in neurons is induced by retinoic acid. The RAI1 gene is associated with both the severity of the phenotype and the response to medication in schizophrenic patients. Defects in RAI1 are a cause of Smith-Magenis syndrome (SMS). There are four named isoforms.
Cellular localizationCytoplasmic and Nuclear. In neurons it is localized to neurites.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling RAI1 with ab86599 at 1/1000 (1µg/ml). Detection: DAB.
ICC/IF image of ab86599 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab86599 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
All lanes : Anti-RAI1 antibody (ab86599) at 0.1 µg/ml
Lane 1 : Whole cell lysate from HeLa cells at 50 µg
Lane 2 : Whole cell lysate from HeLa cells at 15 µg
Lane 3 : Whole cell lysate from HeLa cells at 5 µg
Developed using the ECL technique.
Predicted band size: 203 kDa
Observed band size: 280 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes
Western Blot detection of RAI1 in Immunoprecipitates of Whole cell lysate of HeLa cells (1 mg for IP, 20% of IP loaded) using ab86599 at 10 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection. Lane 2 represents control IgG IP.
Detection: Chemiluminescence with an exposure time of 30 seconds.
ab86599 has not yet been referenced specifically in any publications.