Phospho-MITF (Ser180) Antibody - #AF3027

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Product: Phospho-MITF (Ser180) Antibody
Catalog: AF3027
Description: Rabbit polyclonal antibody to Phospho-MITF (Ser180)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat, Monkey
Prediction: Pig, Bovine, Sheep, Rabbit, Dog, Xenopus
Mol.Wt.: 50~80kDa; 59kD(Calculated).
Uniprot: O75030
RRID: AB_2834458

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Related Downloads
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat,Monkey
Prediction:
Pig(100%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Xenopus(92%)
Clonality:
Polyclonal
Specificity:
Phospho-MITF (Ser180) Antibody detects endogenous levels of MITF only when phosphorylated at Serine 180.
RRID:
AB_2834458
Cite Format: Affinity Biosciences Cat# AF3027, RRID:AB_2834458.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

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

Immunogens

Immunogen:
Uniprot:

O75030(MITF_HUMAN) >>Visit HPA database.

Gene(ID):
MITF(4286)
Expression:
O75030 MITF_HUMAN:

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. Isoform Mdel is widely expressed in melanocytes, melanoma cell lines and tissues, but almost undetectable in non-melanoma cell lines.

Description:
MITF is a transcription factor that contains both basic helix-loop-helix and leucine zipper structural features. Plays a critical role in the differentiation of various cell types including neural crest- derived melanocytes, mast cells, osteoclasts and optic cup-derived retinal pigment epithelium.
Sequence:
MQSESGIVPDFEVGEEFHEEPKTYYELKSQPLKSSSSAEHPGASKPPISSSSMTSRILLRQQLMREQMQEQERREQQQKLQAAQFMQQRVPVSQTPAINVSVPTTLPSATQVPMEVLKVQTHLENPTKYHIQQAQRQQVKQYLSTTLANKHANQVLSLPCPNQPGDHVMPPVPGSSAPNSPMAMLTLNSNCEKEGFYKFEEQNRAESECPGMNTHSRASCMQMDDVIDDIISLESSYNEEILGLMDPALQMANTLPVSGNLIDLYGNQGLPPPGLTISNSCPANLPNIKRELTACIFPTESEARALAKERQKKDNHNLIERRRRFNINDRIKELGTLIPKSNDPDMRWNKGTILKASVDYIRKLQREQQRAKELENRQKKLEHANRHLLLRIQELEMQARAHGLSLIPSTGLCSPDLVNRIIKQEPVLENCSQDLLQHHADLTCTTTLDLTDGTITFNNNLGTGTEANQAYSVPTKMGSKLEDILMDDTLSPVGVTDPLLSSVSPGASKTSSRRSSMSMEETEHTC

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Xenopus
92
Horse
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O75030 As Substrate

Site PTM Type Enzyme
T54 Phosphorylation
S55 Phosphorylation
K79 Sumoylation
K128 Ubiquitination
K140 Sumoylation
S157 Phosphorylation
S180 Phosphorylation P28482 (MAPK1)
K198 Ubiquitination
S216 Phosphorylation
K289 Sumoylation
S301 Phosphorylation
K308 Ubiquitination
R330 Methylation
K350 Ubiquitination
S357 Phosphorylation
Y360 Phosphorylation
K372 Ubiquitination
S405 Phosphorylation P49840 (GSK3A)
S409 Phosphorylation Q15418 (RPS6KA1)
S414 Phosphorylation Q16539 (MAPK14)
K423 Sumoylation
T489 Phosphorylation
S491 Phosphorylation
S502 Phosphorylation
S504 Phosphorylation
S508 Phosphorylation
T510 Phosphorylation
S511 Phosphorylation
S512 Phosphorylation
S515 Phosphorylation
S516 Phosphorylation

Research Backgrounds

Function:

Transcription factor that regulates the expression of genes with essential roles in cell differentiation, proliferation and survival. Binds to M-boxes (5'-TCATGTG-3') and symmetrical DNA sequences (E-boxes) (5'-CACGTG-3') found in the promoters of target genes, such as BCL2 and tyrosinase (TYR). Plays an important role in melanocyte development by regulating the expression of tyrosinase (TYR) and tyrosinase-related protein 1 (TYRP1). Plays a critical role in the differentiation of various cell types, such as neural crest-derived melanocytes, mast cells, osteoclasts and optic cup-derived retinal pigment epithelium.

PTMs:

Phosphorylation at Ser-405 significantly enhances the ability to bind the tyrosinase promoter. Phosphorylated at Ser-180 and Ser-516 following KIT signaling, trigerring a short live activation: Phosphorylation at Ser-180 and Ser-516 by MAPK and RPS6KA1, respectively, activate the transcription factor activity but also promote ubiquitination and subsequent degradation by the proteasome.

Ubiquitinated following phosphorylation at Ser-180, leading to subsequent degradation by the proteasome. Deubiquitinated by USP13, preventing its degradation.

Subcellular Location:

Nucleus.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
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. Isoform Mdel is widely expressed in melanocytes, melanoma cell lines and tissues, but almost undetectable in non-melanoma cell lines.

Subunit Structure:

Homodimer or heterodimer; dimerization is mediated via the coiled coil region. Efficient DNA binding requires dimerization with another bHLH protein. Binds DNA in the form of homodimer or heterodimer with either TFE3, TFEB or TFEC. Interacts with KARS1. Identified in a complex with HINT1 and CTNNB1. Interacts with VSX2 (By similarity).

Family&Domains:

The leucine zipper region is part of a larger coiled coil.

Belongs to the MiT/TFE family.

Research Fields

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

· Human Diseases > Cancers: Specific types > Melanoma.   (View pathway)

· Organismal Systems > Development > Osteoclast differentiation.   (View pathway)

· Organismal Systems > Endocrine system > Melanogenesis.

References

1). TGFβ2 Upregulates Tyrosinase Activity through Opsin-3 in Human Skin Melanocytes In Vitro. Journal of Investigative Dermatology, 2021 (PubMed: 34029574) [IF=6.5]
2). The genetic basis and potential molecular mechanism of yellow-albino northern snakehead (Channa argus). Open Biology, 2023 (PubMed: 36789536) [IF=5.8]

Application: WB    Species: snake    Sample:

Figure 5. The comparison of core melanogenesis genes in skins between WT and YM snakeheads. (a) Western blotting detection of TYR protein levels in skins of WT and YM snakeheads (mean ± s.e.m., n = 4). (b) qPCR detection of mRNA levels of the melanogenic genes (tyr, tyrp1a, tyrp1b, dct and pmel) at different ontogenetic stages (mean ± s.e.m., n = 3). (c) qPCR detection of mRNA levels of mitfa and mitfb in skins of WT and YM snakeheads (mean ± s.e.m., n = 3). (d) Western blotting detection of phospho-MITF(Ser180) protein levels in skins of WT and YM snakeheads (mean ± s.e.m., n = 4). The western blots are cropped and blot source data are available in the electronic supplementary material, figures S3 and S4. * p < 0.05, ** p < 0.01, *** p < 0.001, n.s.: not significant.
3). Cistanche deserticola polysaccharide induces melanogenesis in melanocytes and reduces oxidative stress via activating NRF2/HO-1 pathway. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, 2020 (PubMed: 32096914) [IF=5.3]

Application: WB    Species: human    Sample: HEMs

FIGURE 2|CDP promotes melanogenesis in HEMs. The HEMs were treated with CDP at different concentrations (20, 40 and 80 μg/mL) or medium alone (NC) for 48 h, then we observed the melanin by Fontana-Masson staining, measured the melanin content (OD value, 470 nm) by NaOH assay, and measured the tyrosinase activity (OD value, 475 nm) by tyrosinase activity assay; meanwhile, we measured the protein levels of MITF, p-MITF, TYR, TRP1, TRP2, RAB27A and FSCN1 by Western blotting and measure the grey value of protein bands by Image J: (A) melanin staining; (B) the melanin content; (C) the tyrosinase activity; (D) the protein levels of melanogenesis-related genes
4). p38 MAPK is involved in the immune response to pathogenic Vibrio in the clam Meretrix petechialis. Fish & Shellfish Immunology, 2019 (PubMed: 31669282) [IF=4.7]

Application: WB    Species: clams    Sample: haemocytes

Fig. 8.| (A) Phosphorylated MpMITF (p-MITF) expression in the haemocytes of M. petechialis in the control group and p38 inhibitor group, as measured by Western blot. β-actin was used as the internal reference to normalize the total protein quantity between samples.
5). Exploring the mechanism of Liuwei Dihuang formula for promoting melanin synthesis in juvenile zebrafish based on network pharmacology and molecular docking. Heliyon, 2023 (PubMed: 38027573) [IF=4.0]

Application: WB    Species: zebrafish    Sample:

Fig. 6 LDF promote melanogenesis in juvenile zebrafish. A. Toxicity assays in embryos with drugs administration of LDF (0–120 μg/ml); B. The effect of LDF on melanogenesis with drugs administration of LDF (20–80 μg/ml and 1 μM methoxsalen) for 72 h; C-D. The effect of LDH on melanin contents; E. The effect of LDH on the protein expression of MITF, p-MITF, TYR, TRP1, and TRP2.

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