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Products:Epigenetics and Nuclear Signaling >> Transcription >> Domain Families >> HLH / Leucine Zipper
Overview
Product name
Anti-MiTF antibody
See all MiTF products (10) ...
Description
Rabbit polyclonal to MiTF
Tested applications
ICC/IF, IHC-P, WBmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 500 to the C-terminus of Human MiTF.
(Peptide available as ab21454.)
Positive control
This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) A431 (Human epithelial carcinoma cell line) NIH 3T3 (Mouse embryonic fibroblast cell line) MEF1 (Mouse embryonic fibroblast cell line) PC12 (Rat adrenal pheochromocytoma cell line) This antibody gave a positive signal in HeLa nuclear lysate This antibody gave a positive signal in the following tissue lysates: Brain (Mouse and Rat) Testis (Mouse) Spinal Cord (Mouse) Ovary (Mouse)
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Show applications key
Our Abpromise guarantee covers the use of ab20663 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use at an assay dependent concentration. (PubMed: 22131883)
IHC-P: 1/250Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).Can be blocked with MiTF peptide (ab21454).
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
- Entrez Gene: 17342 Mouse
- Entrez Gene: 25094 Rat
- Omim: 156845 Human
- SwissProt: O75030 Human
- SwissProt: Q08874 Mouse
- SwissProt: O88368 Rat
- Unigene: 166017 Human
see all
Anti-MiTF antibody images:
Western blot - MiTF antibody (ab20663)
All lanes : Anti-MiTF antibody (ab20663) at 1 µg/ml
Lane 1 : HeLa whole cell lysate
Lane 2 : HeLa nuclear lysate
Lane 3 : A431 whole cell lysate
Lane 4 : HeLa whole cell lysate with MiTF peptide (ab21454) at 1 µg/ml
Lane 5 : HeLa nuclear lysate with MiTF peptide (ab21454) at 1 µg/ml
Lane 6 : A431 whole cell lysate with MiTF peptide (ab21454) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution
Predicted band size : 52 kDa
Observed band size : 52 kDa
Additional bands at : 25 kDa,37 kDa. We are unsure as to the identity of these extra bands.
Western blot - MiTF antibody (ab20663)
All lanes : Anti-MiTF antibody (ab20663) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Brain (Mouse) Tissue Lysate
Lane 4 : Testis (Mouse) Tissue Lysate - normal tissue
Lane 5 : Spinal Cord (Mouse) Tissue Lysate
Lane 6 : Ovary (Mouse) Tissue Lysate - normal tissue
Lane 7 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 8 : Brain (Rat) Tissue Lysate - normal tissue
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 52 kDa
Observed band size : 52 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - MiTF antibody (ab20663)
ab20663 staining MiTF in mouse skin tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue section was fixed in formaldehyde and after heat mediated antigen retrieval, blocked (5 minutes/peroxidase block then 10 minutes/protein block) for 15 minutes at 20°C. The primary antibody was diluted, 1/250 and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted, as the secondary.
This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - MiTF antibody (ab20663)
ab20663 staining MiTF in melanoma tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval and then blocking (5 minutes of peroxidase block and 10 minutes of protein block) was performed for 15 minutes at 20°C . The primary antibody was used undiluted and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary antibody.
This image is a courtesy of Antibody Solutions Ltd.
References for Anti-MiTF antibody (ab20663)
This product has been referenced in:
- Katoh Iet al. Activation of the Long Terminal Repeat of Human Endogenous Retrovirus K by Melanoma-Specific Transcription Factor MITF-M. Neoplasia 13:1081-92 (2011). ICC/IF; Human.Read more (PubMed: 22131883) »
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"