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  1. Link

    mitf-antibody-c5-ab12039.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families HLH / Leucine Zipper
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Anti-MiTF antibody [C5] (ab12039)

  • Datasheet
  • SDS
Reviews (5)Q&A (10)References (89)

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Western blot - Anti-MiTF antibody [C5] (ab12039)
  • Flow Cytometry - Anti-MiTF antibody [C5] (ab12039)

Key features and details

  • Mouse monoclonal [C5] to MiTF
  • Suitable for: Flow Cyt, WB
  • Reacts with: Human
  • Isotype: IgG1

You may also be interested in

Protein
Product image
Recombinant Human MiTF protein (ab158898)
Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Primary
Product image
Anti-TRP2/DCT antibody (ab74073)

View more associated products

Overview

  • Product name

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

    Mouse monoclonal [C5] to MiTF
  • Host species

    Mouse
  • Tested applications

    Suitable for: Flow Cyt, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken
  • 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 a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    C5
  • Isotype

    IgG1
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • HLH
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • HLH / Leucine Zipper
    • Immunology
    • Innate Immunity
    • Mast Cells

Associated products

  • Alternative Versions

    • Anti-MiTF antibody [C5] - BSA and Azide free (ab264110)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human MiTF protein (ab158898)

Applications

The Abpromise guarantee

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
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).
Notes
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).

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 accession O75030 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 618266 Human
    • Unigene: 333284 Mouse
    • Unigene: 454504 Mouse
    • Unigene: 31427 Rat
    see all
  • 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

  • Western blot - Anti-MiTF antibody [C5] (ab12039)
    Western blot - Anti-MiTF antibody [C5] (ab12039)
    Anti-MiTF antibody [C5] (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
  • Flow Cytometry - Anti-MiTF antibody [C5] (ab12039)
    Flow Cytometry - Anti-MiTF antibody [C5] (ab12039)
    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.

Protocols

  • Mouse on Mouse staining protocol

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (89)

Publishing research using ab12039? Please let us know so that we can cite the reference in this datasheet.

ab12039 has been referenced in 89 publications.

  • Dilshat R  et al. MITF reprograms the extracellular matrix and focal adhesion in melanoma. Elife 10:N/A (2021). PubMed: 33438577
  • Habel N  et al. FBXO32 links ubiquitination to epigenetic reprograming of melanoma cells. Cell Death Differ 28:1837-1848 (2021). PubMed: 33462405
  • Melixetian M  et al. Long non-coding RNA TINCR suppresses metastatic melanoma dissemination by preventing ATF4 translation. EMBO Rep 22:e50852 (2021). PubMed: 33586907
  • Ouyang Y  et al. UVB-Induced ciRS-7 Activates Melanogenesis by Paracrine Effects. DNA Cell Biol 40:523-531 (2021). PubMed: 33687273
  • Netcharoensirisuk P  et al. Flavonoids increase melanin production and reduce proliferation, migration and invasion of melanoma cells by blocking endolysosomal/melanosomal TPC2. Sci Rep 11:8515 (2021). PubMed: 33875769
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 15 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-MiTF antibody [C5] - ChIP Grade

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 04 2019

Immunohistochemistry (Frozen sections) abreview for Anti-MiTF antibody [C5] - ChIP Grade

Poor
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 24 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-MiTF [C5] antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 16 2014

Immunohistochemistry (Frozen sections) abreview for Anti-MiTF antibody [C5] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Katia Del Rio-Tsomis

Verified customer

Submitted Sep 30 2011

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MiTF antibody [C5] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 26 2011

Question

I'm looking for an MiTF isoform M specific antibody.

Read More

Abcam community

Verified customer

Asked on Dec 20 2012

Answer

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 melanocytes. It is a mouse IgG1 Kappa chain antibody. It is believed to be M form specific, but that has not been specifically validated by Abcam. The antibody was originally made from an N terminal peptide coupled to KLH. I would refer you to the references section of our datasheet.
https://www.abcam.com/mitf-antibody-c5-chip-grade-ab12039.html
https://www.abcam.com/MiTF-antibody-C5-ChIP-Grade-ab12039-references.html
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Dec 20 2012

Question

I have 2 questions concerning Mitf ab 12039 C5 clone:
- What is the specificity of his antibody, does it recognise only Mitf
isoform M or also Mitf Isoforms A, H, and C?
- In the datasheet is written that the immunogen is N-terminal fragment
of human microphthalmia protein. Is this the N-terminal fragment of
different isoforms of Mitf or of only 1 isoform? And if of only 1
isoform, which one was used?

Read More

Abcam community

Verified customer

Asked on Mar 28 2012

Answer

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, but wouldneed to be validated that as the literature was to utilize the antibodies C5 and D5 for diagnostic purposes in melanomas. The antibody was originally made from an N terminal peptide coupled to KLH.

I hope this information is of some help to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Mar 28 2012

Question

I have 2 questions concerning Mitf ab 12039 C5 clone:
- What is the specificity of his antibody, does it recognise only Mitf
isoform M or also Mitf Isoforms A, H, and C?
- In the datasheet is written that the immunogen is N-terminal fragment
of human microphthalmia protein. Is this the N-terminal fragment of
different isoforms of Mitf or of only 1 isoform? And if of only 1
isoform, which one was used?

Read More

Abcam community

Verified customer

Asked on Mar 14 2012

Answer

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.

Read More

Abcam Scientific Support

Answered on Mar 14 2012

Question

I performed all the changes you suggested and am still unable to get a band for MITF using the (C5)mouse monoclonal antibody using an even wider panel of melanoma lines and primary human melanocytes. Once again, I am able to get a band for survivin and beta-actin using the same lysates. Could you send me a replacement antibody for MITF from a different lot of the C5 monoclonal?

Read More

Abcam community

Verified customer

Asked on Aug 29 2006

Answer

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 172077 and will be shipped to you today, Could you please let me know how you get on with the replacement vial?

Read More

Abcam Scientific Support

Answered on Aug 30 2006

Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 166189 DESCRIPTION OF THE PROBLEM Non-specific bands obtained. Specific bands between 52-70 kD not seen. SAMPLE 3 primary melanocyte lines, and 3 melanoma lines lysed in 2% SDS lysis buffer. PRIMARY ANTIBODY MITF (C5)mouse monoclonal antibody by abcam at 1:1000 dilution in 5%milk for 90 minutes, followed by 3 washes (30 minutes) in PBST. As control, beta actin mouse monoclonal (sigma) and survivin rabbit polyclonal (Dr. Altieri) used similarly. DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED The survivin and beta-actin blots as noted above ANTIBODY STORAGE CONDITIONS antibody aliquoted and stored at -20C. Thawed on ice, used at 1:1000 dilution for western. SAMPLE PREPARATION protease and phosphatase inhibitors (sigma) used. Tris used to pH at 7.4. sample heated at 94C for 3 minutes. AMOUNT OF PROTEIN LOADED 30 micrograms ELECTROPHORESIS/GEL CONDITIONS 10% SDS-PAGE TRANSFER AND BLOCKING CONDITIONS 1X Tris-glycine, 20%methanol used as transfer buffer. Transfer carried out on ice 100V for 1 hour. SECONDARY ANTIBODY [another company] goat anti-mouse (and anti-rabbit for survivin) used at 1:10000 in 5%milk. incubated for 30 minutes, followed by 3 washes in PBST for 30 minutes HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? tried lengthening the incubation and wash steps

Read More

Abcam community

Verified customer

Asked on Aug 17 2006

Answer

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 lysis buffer (the recipe can be found, if you need it, on our protocols page on our website) so as to extract optimally the protein, which is in the nucleus (leaving the samples to lyse for 2-4hours in the lysis buffer containing plenty of fresh protease inhibitors) -block the membrane in 5%BSA in TBST (Tween 20 0.1%) for 1 hour, then rinse lightly the membrane 10 seconds in TBST and incubate the antibody in TBST only. -try diluting the antibody more, you may find that with the above changes you can dilute more. If you still experience problems with the above changes please do not hesitate to contact me and I can arrange for a replacement vial or refund as you purchased the antibody in the last 90 days,

Read More

Abcam Scientific Support

Answered on Aug 17 2006

1-10 of 15 Abreviews or Q&A

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