Product nameAnti-MiTF antibody [C5] - ChIP Grade
See all MiTF primary antibodies
DescriptionMouse monoclonal [C5] to MiTF - ChIP Grade
SpecificityThe antibody recognizes serine phosphorylated and non-phosphorylated melanocytic isoforms of microphthalmia.
Tested applicationsSuitable for: IHC-Fr, IHC-P, ICC/IF, ChIP, Flow Cyt, WB, IP, GSAmore details
Species reactivityReacts with: Mouse, Rat, Chicken, Human
Recombinant fragment corresponding to Human MiTF (N terminal).
- WB: Malme-3M whole cell lysate. Flow Cyt: Malme-3M cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
- Epigenetics and Nuclear Signaling
- Domain Families
- HLH / Leucine Zipper
- HLH / Leucine Zipper
ChIP Related Products
Our Abpromise guarantee covers the use of ab12039 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 µg/ml. Antigen retrieval is not essential but may optimise staining.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 16648630
Fix with methanol/acetone, block with 1% BSA/PBS (see Wellbrock et al).
|ChIP||Use at an assay dependent concentration. PubMed: 21990371|
|EMSA||Use at an assay dependent concentration.|
|Flow Cyt||Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 52-56 kDa (predicted molecular weight: 59 kDa).|
|IP||Use at 2 µg/mg of lysate.|
|GSA||Use at an assay dependent concentration.|
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
Anti-MiTF antibody [C5] - ChIP Grade (ab12039) at 1 µg/ml + Malme 3m (Human melanoma cells) Whole Cell Lysate at 10 µg
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: 40,55,70 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutes
ab12039 staining MiTF in embryonic day 4 chick eye tissue by Immunohistochemistry (Frozen sections).
Tissue was fixed with 4% paraformaldehyde, permeabilized using 1% saponin, blocked with 10% serum for 30 minutes at 37°C, then incubated with ab12039 at a 1/1000 dilution for 24 hours at 4deg;C. The secondary used was an Alexa-Fluor 488 conjugated mouse monoclonal, used at a 1/100 dilution. Counterstained with DAPI (blue).
Overlay histogram showing Malme 3m cells stained with ab12039 (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 followed by the antibody (ab12039, 2µg/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:
- Chiu CS et al. Exploiting Honokiol-induced ER stress CHOP activation inhibits the growth and metastasis of melanoma by suppressing the MITF and ß-catenin pathways. Cancer Lett 442:113-125 (2019). Read more (PubMed: 30391358) »
- Yun CY et al. Nuclear Entry of CRTC1 as Druggable Target of Acquired Pigmentary Disorder. Theranostics 9:646-660 (2019). Read more (PubMed: 30809299) »