Anti-MiTF antibody [D5] (ab3201)

Overview

• Product nameAnti-MiTF antibody [D5]See all MiTF primary antibodies ...
• Description
  Mouse 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 reactivity
  Reacts with: Dog, Human
  Predicted to work with: Mouse

  Does not react with

  Rat
• Immunogen

  N-terminal fragment of human Mi protein.

• EpitopeN-terminal
• Positive control501 Mel cells Melanoma
  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).

Properties

• FormLiquid
• 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
• IsotypeIgG1
• Light chain typekappa
• Research Areas
  • Epigenetics and Nuclear Signaling
  • Transcription
  • Domain Families
  • HLH / Leucine Zipper
  • HLH / Leucine Zipper

  • Immunology
  • Innate Immunity
  • Mast Cells

Applications

Target

• 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.
• Post-translational
  modificationsPhosphorylation at Ser-405 significantly enhances the ability to bind the tyrosinase promoter.
• Cellular localizationNucleus.

Target information above from: UniProt accession O75030 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• 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

  • Unigene: 454504 Mouse

  see all

• Alternative names
  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

  see all

Anti-MiTF antibody [D5] images

• 

  Western blot - 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
  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)
  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)This image is courtesy of an anonymous Abreview
  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.

  See Abreview