Overview

  • Product name

    Anti-MiTF antibody [D5] - BSA and Azide free
    See all MiTF primary antibodies
  • Description

    Mouse monoclonal [D5] to MiTF - BSA and Azide free
  • Host species

    Mouse
  • Specificity

    ab3201 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 applications

    Suitable for: IHC-P, GSA, Flow Cyt, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Dog, Human
    Predicted to work with: MouseDoes not react with: Rat
  • Immunogen

    corresponding to MiTF.

  • Epitope

    N-terminal
  • Positive control

    • FC: Malme-3 cells. ICC/IF: Malme-3 cells. WB: HeLa whole cell and nuclear lysate.
  • General notes

    ab182842 is the carrier-free version of ab3201. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab182842 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab182842 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-P Use at an assay dependent concentration.
GSA Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.
WB Use at an assay dependent concentration. Predicted molecular weight: 59 kDa.
ICC/IF Use at an assay dependent concentration.
EMSA Use a concentration of 1 mg/ml.

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

  • 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab3201).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab3201).

References

ab182842 has not yet been referenced specifically in any publications.

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