- Datasheet
- Specific References (7)
- Protocols
Overview
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Product nameAnti-MiTF antibody [D5]
See all MiTF primary antibodies -
DescriptionMouse monoclonal [D5] to MiTF
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Host speciesMouse
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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.
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Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
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Species reactivityReacts with: Dog, Human
Predicted to work with: Mouse
Does not react with:
Rat -
Immunogen
N-terminal fragment of human Mi protein.
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EpitopeN-terminal
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Positive control
- 501 Mel cells
Melanoma
In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised Malme-3M cells.
- 501 Mel cells
Melanoma
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General notes
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading... -
PurityIgG fraction
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ClonalityMonoclonal
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Clone numberD5
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IsotypeIgG1
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Light chain typekappa
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| EMSA | Use at an assay dependent concentration. Use at an assay dependent concentration |
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| ICC/IF | Use at an assay dependent concentration. | |
| WB | Use at an assay dependent concentration. Predicted molecular weight: 59 kDa. | |
| IHC-P | Use at an assay dependent concentration. PubMed: 18638353 | |
| Flow Cyt | Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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Target
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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.
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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.
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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. -
Post-translational
modificationsPhosphorylation at Ser-405 significantly enhances the ability to bind the tyrosinase promoter. -
Cellular localizationNucleus.
- Information by UniProt
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Database links
- Entrez Gene: 4286 Human
- Entrez Gene: 17342 Mouse
- Omim: 156845 Human
- SwissProt: O75030 Human
- SwissProt: Q08874 Mouse
- Unigene: 166017 Human
- Unigene: 618266 Human
- Unigene: 333284 Mouse
see all -
Alternative names
- BHLHE32 antibody
- bHLHe32 antibody
- Class E basic helix-loop-helix protein 32 antibody
see all
Images
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![Immunocytochemistry/ Immunofluorescence - Anti-MiTF antibody [D5] (ab3201)](https://www.abcam.com/ps/products/3/ab3201/Images/ab3201-234149-ap835815edited.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-MiTF antibody [D5] (ab3201)ICC/IF image of ab3201 stained Malme-3M 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 ab3201 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- mouse (ab150117) 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.
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![Western blot - Anti-MiTF antibody [D5] (ab3201)](https://www.abcam.com/ps/products/3/ab3201/Images/MiTF-Primary-antibodies-ab3201-4.jpg)
Western blot - Anti-MiTF antibody [D5] (ab3201)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
All lanes : 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)](https://www.abcam.com/ps/products/3/ab3201/Images/MiTF-Primary-antibodies-ab3201-6.jpg)
Flow Cytometry - Anti-MiTF antibody [D5] (ab3201)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. -
![Immunocytochemistry/ Immunofluorescence - Anti-MiTF antibody [D5] (ab3201)](https://www.abcam.com/ps/products/3/ab3201/Images/MiTF-Primary-antibodies-ab3201-14.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-MiTF antibody [D5] (ab3201)This image is courtesy of an anonymous Abreviewab3201 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.
Protocols
References
This product has been referenced in:
- Saini JS et al. Nicotinamide Ameliorates Disease Phenotypes in a Human iPSC Model of Age-Related Macular Degeneration. Cell Stem Cell : (2017). Read more (PubMed: 28132833) »
- Herms A et al. AMPK activation promotes lipid droplet dispersion on detyrosinated microtubules to increase mitochondrial fatty acid oxidation. Nat Commun 6:7176 (2015). WB . Read more (PubMed: 26013497) »
Customer reviews and Q&As
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