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 applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
Species reactivityReacts with: Dog, Human
Predicted to work with: MouseDoes not react with: Rat
N-terminal fragment of human Mi protein.
- 501 Mel cells
In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised Malme-3M cells.
- 501 Mel cells Melanoma
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: PBS, 6.97% L-Arginine
Concentration information loading...
Light chain typekappa
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.
|EMSA||Use at an assay dependent concentration.
Use at an assay dependent concentration
|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.
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.
- Information by UniProt
- BHLHE32 antibody
- bHLHe32 antibody
- Class E basic helix-loop-helix protein 32 antibody
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.
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
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.
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.
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.
This product has been referenced in:
- Fernandes M et al. Stem Cell-Derived Retinal Pigment Epithelial Layer Model from Adult Human Globes Donated for Corneal Transplants. Curr Protoc Stem Cell Biol 45:e53 (2018). Read more (PubMed: 30040247) »
- 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) »