Overview

  • Product name

    Anti-MiTF antibody [C5] - ChIP Grade
    See all MiTF primary antibodies
  • Description

    Mouse monoclonal [C5] to MiTF - ChIP Grade
  • Host species

    Mouse
  • Specificity

    The antibody recognizes serine phosphorylated and non-phosphorylated melanocytic isoforms of microphthalmia.
  • Tested applications

    Suitable for: IHC-Fr, IHC-P, ICC/IF, ChIP, Flow Cyt, WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Chicken, Human
  • Immunogen

    Recombinant fragment corresponding to Human MiTF (N terminal).

  • Positive control

    • WB: Malme-3M whole cell lysate. Flow Cyt: Malme-3M cells.
  • 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

Applications

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.

Application Abreviews Notes
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.

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.
  • Information by UniProt
  • Database links

  • Alternative names

    • BHLHE32 antibody
    • bHLHe32 antibody
    • Class E basic helix-loop-helix protein 32 antibody
    • CMM8 antibody
    • Homolog of mouse microphthalmia antibody
    • Mi antibody
    • Microphthalmia associated transcription factor antibody
    • Microphthalmia, mouse, homolog of antibody
    • Microphthalmia-associated transcription factor antibody
    • MITF antibody
    • MITF_HUMAN antibody
    • mitfa antibody
    • nacre antibody
    • WS2 antibody
    • WS2A antibody
    • z3A.1 antibody
    see all

Images

  • Anti-MiTF antibody [C5] - ChIP Grade (ab12039) at 1 µg/ml + Malme 3m (Human melanoma cells) Whole Cell Lysate at 10 µg

    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: 40,55,70 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).

    See Abreview

  • 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.

References

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) »
See all 54 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

1-5 of 5 Abreviews

Application
Immunohistochemistry (Frozen sections)
Sample
Chicken Tissue sections (Cryostat tissue sections of 40um, from neural cres)
Permeabilization
Yes - 0.5% triton x-100
Specification
Cryostat tissue sections of 40um, from neural cres
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 04 2019

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Sample
Mouse Tissue sections (e11.5 embryo transverse cryosection of trunk)
Specification
e11.5 embryo transverse cryosection of trunk
Permeabilization
Yes - 1% saponin
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 24 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
2.5% goat serum, 2.5% donkey serum, 1% BSA, 0.1% triton as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Sample
Human Cell (neonatal foreskin melanocytes)
Specification
neonatal foreskin melanocytes
Permeabilization
Yes - 0.1% tritonx-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 16 2014

Application
Immunohistochemistry (Frozen sections)
Sample
Chicken Tissue sections (eye)
Specification
eye
Fixative
Paraformaldehyde
Permeabilization
Yes - 1% saponin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C

Katia Del Rio-Tsomis

Verified customer

Submitted Sep 30 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin)
Specification
skin
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric buffer, pH6
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted May 26 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up