Overview

  • Product name
    Anti-MelanA antibody [EP1422Y]
    See all MelanA primary antibodies
  • Description
    Rabbit monoclonal [EP1422Y] to MelanA
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, IHC-Fr, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human MelanA aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: Q16655

  • Positive control
    • Melanoma lysate or melanoma tissue.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab51061 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
WB 1/1000 - 1/10000. Detects a band of approximately 20 kDa (predicted molecular weight: 13 kDa).
IHC-P 1/100 - 1/250.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Involved in melanosome biogenesis by ensuring the stability of GPR143. Plays a vital role in the expression, stability, trafficking, and processing of melanocyte protein PMEL, which is critical to the formation of stage II melanosomes.
    • Tissue specificity
      Expression is restricted to melanoma and melanocyte cell lines and retina.
    • Post-translational
      modifications
      Acylated.
    • Cellular localization
      Endoplasmic reticulum membrane. Golgi apparatus. Golgi apparatus > trans-Golgi network membrane. Melanosome. Also found in small vesicles and tubules dispersed over the entire cytoplasm. A small fraction of the protein is inserted into the membrane in an inverted orientation. Inversion of membrane topology results in the relocalization of the protein from a predominant Golgi/post-Golgi area to the endoplasmic reticulum. Melanoma cells expressing the protein with an inverted membrane topology are more effectively recognized by specific cytolytic T-lymphocytes than those expressing the protein in its native membrane orientation.
    • Information by UniProt
    • Database links
    • Alternative names
      • Antigen LB39 AA antibody
      • Antigen LB39-AA antibody
      • Antigen SK29 AA antibody
      • Antigen SK29-AA antibody
      • MAR1_HUMAN antibody
      • MART 1 antibody
      • MART-1 antibody
      • MART1 antibody
      • Melan A antibody
      • Melan A protein antibody
      • Melanoma antigen recognized by T cells 1 antibody
      • Melanoma antigen recognized by T-cells 1 antibody
      • MLAN A antibody
      • MLANA antibody
      • OTTHUMP00000021036 antibody
      • OTTHUMP00000021037 antibody
      • OTTHUMP00000021038 antibody
      • Protein Melan-A antibody
      see all

    Images

    • Ab51061 (1/100) staining human MelanA in melanoma tissue by immunohistochemistry using paraffin embedded tissue.
    • Immunohistochemical analysis of Human malignant melanoma, staining MelanA with ab51061 at 1/20000 dilution. A Cy3-conjugated donkey anti-rabbit IgG (1/500) was used as the secondary antibody.
    • Overlay histogram showing Malme-3M cells stained with ab51061 (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 (ab51061, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Anti-MelanA antibody [EP1422Y] (ab51061) at 1/1000 dilution + Melanoma lysate at 10 µg

      Secondary
      Goat anti-Rabbit HRP labeled at 1/2000 dilution

      Predicted band size: 13 kDa
      Observed band size: 20 kDa
      why is the actual band size different from the predicted?

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:
    • Regazzetti C  et al. Melanocytes Sense Blue Light and Regulate Pigmentation through Opsin-3. J Invest Dermatol 138:171-178 (2018). Read more (PubMed: 28842328) »
    • Patwardhan A  et al. Routing of the RAB6 secretory pathway towards the lysosome related organelle of melanocytes. Nat Commun 8:15835 (2017). ICC/IF . Read more (PubMed: 28607494) »
    See all 15 Publications for this product

    Customer reviews and Q&As

    1-5 of 5 Abreviews or Q&A

    Question
    Answer

    Merci de nous avoir contactés.

    Suite à notre conversation téléphonique, je me suis renseignée a propos de vos recherches. Apres réflexion, j’ai du mal à comprendre comment vous allez pouvoir faire de l’immunofluorescence en vivo chez l’humain puisque vous allez être obligé de couper les grains de beauté afin de les regarder sous le microscope confocal donc, cette expérience ne serra pas en vivo. Nous aimerions vous rappeler que nos produits sont pour la recherche uniquement et ne sont pas destiné à être utilisés pour des fins diagnostiques ou thérapeutiques.

    A mon avis, cette expérience ne donnera pas de résultats positifs à moins de faire des coupes de grain de beauté. Si vous ne faites pas de coupes, le grain de beauté sera trop épais pour que la lumière du microscope confocal passe à travers et détecte le fluorophore.

    Je vous suggère donc d’essayer de l’immunohistochimie sur des coupes de paraffines ou sur des coupes congèles. Pour préparer des coupes de paraffines, vous devez dans un premier temps fixer le tissue dans 4% PFA puis le mettre dans de la paraffine avant de faire vos coupes.

    Pour préparer des coupes congèles, vous devez premièrement cryo-congeler votre tissue et faire des coupes puis les fixer.

    Je vous attache quelques protocoles que l’on suggère.

    Je peux vous proposer les anticorps suivant pour l’immunohistochimie :

    “Anti-MelanA antibody [EP1422Y] (ab51061)” (https://www.abcam.com/ab51061) avec comme anticorps secondaire “Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)” (ab150077)(https://www.abcam.com/ab150077)

    Ou

    “Anti-MelanA antibody [DT101 + BC199] (ab731)”(https://www.abcam.com/ab731) avec comme anticorps secondaire “Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 488) (ab150113)”(https://www.abcam.com/ab150113)

    J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

    Monoclonal de lapin gratuit pour tout achat d'anticorps primaire, dans la limite des stocks disponibles. Notez le code "RABMAB-XBSMG" sur votre prochaine commande d’anticorps primaire. Pour plus d'informations, visitez le lien suivant: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Answer

    Thank you for your enquiry.

    Indeed, the purification kits are designed for cleaning up small volumes of antibodies (ideallyin a volume of 100µl to 0.5ml), as you can see on the datasheet for ab102784.

    https://www.abcam.com/index.html?datasheet=102784 (or use the following: https://www.abcam.com/index.html?datasheet=102784).

    The only issue to consider with these purification kitsis thatthey arenot suitable for goat, sheep or rat antibodies due to the low affinity for Protein A. However, the MelanA antibodies you are interested in are raised in rabbit, so this is not a problem here at all.

    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

    Answer

    Thank you for contacting us.

    Having checked with the lab, I am sorry to let you know that we are unable to provide these MelanA antibodies in a buffer without BSA.

    However, if you need to remove BSA and other unwanted ingredients, I may recommend using one of our purification kits such as ab102783 which are designed for this purpose. Further information can be found on our website at https://www.abcam.com/index.html?pageconfig=resource&rid=13489.

    ab102783: https://www.abcam.com/index.html?datasheet=102783 (or use the following: https://www.abcam.com/index.html?datasheet=102783).

    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

    Question
    Answer

    You are welcome! We are always happy to help :-) Have a great day!

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (skin, melanozytes, 10ym, OCT embedded)
    Specification
    skin, melanozytes, 10ym, OCT embedded
    Fixative
    Methanol
    Permeabilization
    No

    Dr. Christian Posch

    Verified customer

    Submitted Oct 21 2011

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

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