Anti-MiTF抗体[C5] - ChIP Grade (ab12039)

概述

  • 产品名称
    Anti-MiTF抗体[C5] - ChIP Grade
    参阅全部 MiTF 一抗
  • 描述
    小鼠单克隆抗体[C5] to MiTF - ChIP Grade
  • 特异性
    The antibody recognizes serine phosphorylated and non-phosphorylated melanocytic isoforms of microphthalmia.
  • 经测试应用
    适用于: IHC-Fr, IHC-P, ICC/IF, ChIP, Flow Cyt, WB, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Human
  • 免疫原

    Recombinant fragment corresponding to Human MiTF (N terminal).

  • 阳性对照
    • This antibody gave a positive signal in Malme-3M whole cell lysate. In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised Malme-3M cells.
  • 常规说明

    This antibody clone is manufactured by Abcam.

    We can conjugate this antibody to FITC for you (please see ab150195 for details). Do not store in a frost-free freezer. The antibody is stable for at least 12 months when stored appropriately. Working dilutions should be discarded if not used within 12 hours.

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能
    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.
  • 组织特异性
    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.
  • 疾病相关
    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.
  • 序列相似性
    Belongs to the MiT/TFE family.
    Contains 1 basic helix-loop-helix (bHLH) domain.
  • 翻译后修饰
    Phosphorylation at Ser-405 significantly enhances the ability to bind the tyrosinase promoter.
  • 细胞定位
    Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-MiTF antibody [C5] - ChIP Grade 图像

  • 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.

Anti-MiTF antibody [C5] - ChIP Grade (ab12039)参考文献

This product has been referenced in:
  • Schwochow Thalmann D  et al. The evolution of Sex-linked barring alleles in chickens involves both regulatory and coding changes in CDKN2A. PLoS Genet 13:e1006665 (2017). IHC-Fr ; Chicken . Read more (PubMed: 28388616) »
  • Wang X  et al. Efficient CRISPR/Cas9-mediated biallelic gene disruption and site-specific knockin after rapid selection of highly active sgRNAs in pigs. Sci Rep 5:13348 (2015). WB ; Pig . Read more (PubMed: 26293209) »

See all 27 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 Apr 16 2014

Thank you for contacting us.
We carry ab12039 and this antibody recognizes serine phosphorylated and non-phosphorylated melanocytic isoforms of microphthalmia. The Mitf antibody ab12039 [C5] recognizes Mouse, Rat, Chicken, Human MiTF in melanocyte...

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Thank you for your patience.

The following information was obtained:

The Mitf antibody ab12039 [C5] recognizes both human and mouse Mitf in melanocytes. It is a mouse IgG1 Kappa chain antibody. It isbelieved to beM form specific, b...

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Thank you for contacting us.

I have been in touch with the lab. We need to check with the originator of the antibody for these details. I will let you know as soon asI hear back.

Thank you for your patience.

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Katia Del Rio-Tsomis

Verified customer

提交于 Sep 30 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 May 26 2011

Thank you for your e-mail and informing me you have tried the suggested modifications to your protocol, I appreciate the amount of work you have performed and have immediately arranged a replacement vial to be sent to you. The order is reference 172...

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I'm very sorry to hear you are experiencing problems with ab12039 and thank you for providing your protocol details so we can look into your problem. I would like to recommend the following modifications to your protocol: -use a RIPA or NP40 lysi...

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Thank you for getting back to me. I appreciate the steps that you have taken to try and get this antibody to work. I have checked our records and we do not have any other complaints with regards this batch. I would be happy to provide you with a...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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