![Western blot - MiTF antibody [D5] (ab3201) image](#MiTF-Primary-antibodies-ab3201-4.jpg)
 image](#MiTF-Primary-antibodies-ab3201-6.jpg)
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Products:Epigenetics and Nuclear Signaling >> Transcription >> Domain Families >> HLH / Leucine Zipper
Overview
Product name
Anti-MiTF antibody [D5]
See all MiTF products (10) ...
Description
Mouse monoclonal [D5] to MiTF
Specificity
ab3201 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 applications
WB, IHC-P, Flow Cyt, Gel supershift assaysmore details
Cross reactivity
Reacts with
Dog, Human
Does not react with
Mouse, Rat
Immunogen
N-terminal fragment of human Mi protein.
Epitope
N-terminal
Positive control
501 Mel cells Melanoma
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
10mM PBS, pH7.4, 0.2%BSA, 0.09% sodium azide
Concentration
Concentration information loading...
Purity
IgG fraction
Clonality
Monoclonal
Clone number
D5
Isotype
IgG1
Light chain type
kappa
Applications
Show applications key
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.
WB: Use at an assay dependent dilution. Predicted molecular weight: 59 kDa.
IHC-P: Use at an assay dependent dilution.
Flow Cyt: Use 1µg for 106 cells.
GSA: Use a concentration of 1 mg/ml
Target
Function
Transcription 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 specificity
Isoform 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 disease
Defects 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 similarities
Belongs to the MiT/TFE family.
Contains 1 basic helix-loop-helix (bHLH) domain.
Post-translational
modifications
Phosphorylation at Ser-405 significantly enhances the ability to bind the tyrosinase promoter.
Cellular localization
Nucleus.
Target information above from: UniProt accessionO75030
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- BHLHE32 antibody
- bHLHe32 antibody
- Class E basic helix-loop-helix protein 32 antibody
see all
Database links
- Entrez Gene: 4286 Human
- Omim: 156845 Human
- SwissProt: A1E097 Dog
- SwissProt: O75030 Human
- Unigene: 166017 Human
- Unigene: 618266 Human
Anti-MiTF antibody [D5] images:
Western blot - MiTF antibody [D5] (ab3201)
![Western blot - MiTF antibody [D5] (ab3201)](/ps/datasheet/images/3/ab3201/MiTF-Primary-antibodies-ab3201-4.jpg)
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.
Secondary
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
Flow Cytometry-Anti-MiTF antibody [D5](ab3201)
](/ps/datasheet/images/3/ab3201/MiTF-Primary-antibodies-ab3201-6.jpg)
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.
References for Anti-MiTF antibody [D5] (ab3201)
This product has been referenced in:
- Petersen-Jones SMet 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) »
See 1 publication for this product
Publishing research using ab3201? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"