Anti-MiTF antibody [D5] (ab3201)
- Product nameAnti-MiTF antibody [D5]See all MiTF primary antibodies ...
- DescriptionMouse monoclonal [D5] to MiTF
- Specificityab3201 does not cross-react with other b-HLH-ZIP factors by DNA mobility shift assay. It reacts with both melanocytic and non-melanocytic isoforms of Mi. It is especially useful for research on murine Mi protein.
- Tested applicationsICC/IF, WB, IHC-P, Flow Cyt, Gel supershift assays more details
- Species reactivityReacts with: Dog, Human
Predicted to work with: Mouse
Does not react withRat
N-terminal fragment of human Mi protein.
- Positive control501 Mel cells
In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised Malme-3M cells.
- General notesWe can conjugate this antibody to FITC for you (please see ab150228 for details).
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage buffer10mM PBS, pH7.4, 0.2%BSA, 0.09% sodium azide
- Concentration information loading...
- PurityIgG fraction
- Clonality Monoclonal
- Clone numberD5
- Light chain typekappa
- Research Areas
Our Abpromise guarantee covers the use of ab3201 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||ICC/IF: Use at an assay dependent concentration.|
|WB||WB: Use at an assay dependent dilution. Predicted molecular weight: 59 kDa.|
|IHC-P||IHC-P: Use at an assay dependent dilution.|
|Flow Cyt||Flow Cyt: Use 1µg for 106 cells.|
|Gel supershift assays||GSA: Use a concentration of 1 mg/ml.|
- FunctionTranscription factor for tyrosinase and tyrosinase-related protein 1. Binds to a symmetrical DNA sequence (E-boxes) (5'-CACGTG-3') found in the tyrosinase promoter. Plays a critical role in the differentiation of various cell types as neural crest-derived melanocytes, mast cells, osteoclasts and optic cup-derived retinal pigment epithelium.
- Tissue specificityIsoform M is exclusively expressed in melanocytes and melanoma cells. Isoform A and isoform H are widely expressed in many cell types including melanocytes and retinal pigment epithelium (RPE). Isoform C is expressed in many cell types including RPE but not in melanocyte-lineage cells.
- Involvement in diseaseDefects in MITF are the cause of Waardenburg syndrome type 2A (WS2A) [MIM:193510]. It is a dominant inherited disorder characterized by sensorineural hearing loss and patches of depigmentation. The features show variable expression and penetrance.
Defects in MITF are a cause of Waardenburg syndrome type 2 with ocular albinism (WS2-OA) [MIM:103470]. It is an ocular albinism with sensorineural deafness.
Defects in MITF are the cause of Tietz syndrome (TIETZS) [MIM:103500]. It is an autosomal dominant disorder characterized by generalized hypopigmentation and profound, congenital, bilateral deafness. Penetrance is complete.
- Sequence similaritiesBelongs to the MiT/TFE family.
Contains 1 basic helix-loop-helix (bHLH) domain.
modificationsPhosphorylation at Ser-405 significantly enhances the ability to bind the tyrosinase promoter.
- Cellular localizationNucleus.
- BHLHE32 antibodybHLHe32 antibodyClass E basic helix-loop-helix protein 32 antibody
- CMM8 antibodyHomolog of mouse microphthalmia antibodyMi antibodyMicrophthalmia associated transcription factor antibodyMicrophthalmia, mouse, homolog of antibodyMicrophthalmia-associated transcription factor antibodyMITF antibodyMITF_HUMAN antibodyWS2 antibodyWS2A antibody
Anti-MiTF antibody [D5] images
All lanes : Anti-MiTF antibody [D5] (ab3201) at 5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lysates/proteins at 10 µg per lane.
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 59 kDa
Observed band size : 40 kDa (why is the actual band size different from the predicted?)
Exposure time : 10 minutes
Overlay histogram showing Malme-3 cells stained with ab3201 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3201, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.
ab3201 staining MiTF in the SK-MEL-28 (a human melanoma cell line) by ICC/IF(Immunocytochemistry/immunofluorescence). Cells were fixed with Paraformaldehyde and permeabilized with Triton X-100 0.5% in PBS. Samples were incubated with primary antibody (1/100) for 24 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal(1/1000) was used as the secondary antibody. SK-MEL-28 cells was grown on chamber slide. MiTF was found to strictly localised in the nucleus. Cells were counter-stained with DAPI.
References for Anti-MiTF antibody [D5] (ab3201)
This product has been referenced in:
- Zahabi A et al. A New Efficient Protocol for Directed Differentiation of Retinal Pigmented Epithelial Cells from Normal and Retinal Disease Induced Pluripotent Stem Cells. Stem Cells Dev : (2012). Read more (PubMed: 22145677) »
- Petersen-Jones SM et al. Ocular melanosis in the Cairn Terrier: histopathological description of the condition, and immunohistological and ultrastructural characterization of the characteristic pigment-laden cells. Vet Ophthalmol 11:260-8 (2008). IHC-P ; Dog . Read more (PubMed: 18638353) »