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重组RabMAb

重组Anti-MelanA抗体[EP1422Y] (ab51061)

Reviews (1)Q&A (4)References (27)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EP1422Y] (ab51061)
  • Immunocytochemistry/ Immunofluorescence - Anti-MelanA antibody [EP1422Y] (ab51061)
  • Western blot - Anti-MelanA antibody [EP1422Y] (ab51061)
  • Western blot - Anti-MelanA antibody [EP1422Y] (ab51061)
  • Flow Cytometry (Intracellular) - Anti-MelanA antibody [EP1422Y] (ab51061)
  • Flow Cytometry (Intracellular) - Anti-MelanA antibody [EP1422Y] (ab51061)
  • OI-RD Scanning - Anti-MelanA antibody [EP1422Y] (ab51061)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EP1422Y] (ab51061)
  • Anti-MelanA antibody [EP1422Y] (ab51061)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1422Y] to MelanA
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase APC Carrier Free HRP PE

概述

  • 产品名称

    Anti-MelanA抗体[EP1422Y]
    参阅全部 MelanA 一抗

  • 描述

    兔单克隆抗体[EP1422Y] to MelanA

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details

  • 种属反应性

    与反应: Human

  • 免疫原

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

  • 阳性对照

    • WB: Human melanoma lysates. IHC-P: Human melanoma tissue. Flow Cyt (intra): A375 and Malme-3M cells. ICC/IF: MeWo cells.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab51061于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt (Intra)
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/20000. Detects a band of approximately 20 kDa (predicted molecular weight: 13 kDa).
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
ICC/IF
1/100.
说明
Flow Cyt (Intra)
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/20000. Detects a band of approximately 20 kDa (predicted molecular weight: 13 kDa).
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
ICC/IF
1/100.

靶标

  • 功能

    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.

  • 组织特异性

    Expression is restricted to melanoma and melanocyte cell lines and retina.

  • 翻译后修饰

    Acylated.

  • 细胞定位

    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

  • 数据库链接

    • 别名

      • 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

    图片

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EP1422Y] (ab51061)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EP1422Y] (ab51061)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue sections labeling MelanA with purified ab51061 at 1/1000 dilution (1.15 µg/mL). Heat mediated antigen retrieval using Bond™™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    • Immunocytochemistry/ Immunofluorescence - Anti-MelanA antibody [EP1422Y] (ab51061)
      Immunocytochemistry/ Immunofluorescence - Anti-MelanA antibody [EP1422Y] (ab51061)
      Immunocytochemistry/ Immunofluorescence analysis of MeWo (Human malignant melanoma fibroblast) cells labeling MelanA with Purified ab51061 at 1/100 dilution (10 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Western blot - Anti-MelanA antibody [EP1422Y] (ab51061)
      Western blot - Anti-MelanA antibody [EP1422Y] (ab51061)
      Anti-MelanA antibody [EP1422Y] (ab51061) at 1/20000 dilution + MeWo (Human malignant melanoma fibroblast) whole cell lysate at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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



      Blocking buffer: 5% NFDM/TBST.

    • Western blot - Anti-MelanA antibody [EP1422Y] (ab51061)
      Western blot - Anti-MelanA antibody [EP1422Y] (ab51061)
      Anti-MelanA antibody [EP1422Y] (ab51061) at 1/20000 dilution (Purified) + Human melanoma lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

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



      Blocking/Diluting buffer: 5% NFDM/TBST

    • Flow Cytometry (Intracellular) - Anti-MelanA antibody [EP1422Y] (ab51061)
      Flow Cytometry (Intracellular) - Anti-MelanA antibody [EP1422Y] (ab51061)

      Intracellular Flow Cytometry analysis of A375 (Human malignant melanoma epithelial cell, Left) / MeWo (Human malignant melanoma fibroblast, Right) cells labeling MelanA with Purified ab51061 at 1/1000 dilution (1 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

       

    • Flow Cytometry (Intracellular) - Anti-MelanA antibody [EP1422Y] (ab51061)
      Flow Cytometry (Intracellular) - Anti-MelanA antibody [EP1422Y] (ab51061)

      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. 

      This image was generated using the un-purified format of the antibody. 

       

       

    • OI-RD Scanning - Anti-MelanA antibody [EP1422Y] (ab51061)
      OI-RD Scanning - Anti-MelanA antibody [EP1422Y] (ab51061)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EP1422Y] (ab51061)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EP1422Y] (ab51061)

      Tissue Microarrays stained for "Anti-MelanA antibody [EP1422Y]” using "ab51061" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab51061 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Anti-MelanA antibody [EP1422Y] (ab51061)
      Anti-MelanA antibody [EP1422Y] (ab51061)

    实验方案

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    文献 (27)

    发表研究结果有使用 ab51061？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab51061 被引用在 27 文献中.

    • Jung SY  et al. Wnt-activating human skin organoid model of atopic dermatitis induced by Staphylococcus aureus and its protective effects by Cutibacterium acnes. iScience 25:105150 (2022). PubMed: 36193049
    • Beasley AB  et al. Multi-Marker Immunomagnetic Enrichment of Circulating Melanoma Cells. Methods Mol Biol 2265:213-222 (2021). PubMed: 33704717
    • Romo-Valera C  et al. Characterisation of corneas following different time and storage methods for their use as a source of stem-like limbal epithelial cells. Exp Eye Res 211:108720 (2021). PubMed: 34389315
    • Rosenbaum SR  et al. FOXD3 Regulates VISTA Expression in Melanoma. Cell Rep 30:510-524.e6 (2020). PubMed: 31940493
    • Tsering T  et al. Uveal Melanoma-Derived Extracellular Vesicles Display Transforming Potential and Carry Protein Cargo Involved in Metastatic Niche Preparation. Cancers (Basel) 12:N/A (2020). PubMed: 33050649
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-5 of 5 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-MelanA antibody [EP1422Y]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (skin, melanozytes, 10ym, OCT embedded)
    Specification
    skin, melanozytes, 10ym, OCT embedded
    Fixative
    Methanol
    Permeabilization
    No
    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.

    DR. Christian Posch

    Verified customer

    提交于 Oct 21 2011

    Question

    un anticorps anti-melanA et un anticorps secondaire conjugé avec un fluorophore 488.

    Read More
    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

    Question

    thank you very much for your reply. However, it is unfortunate that you cannot provide the antibodies without BSA.
    Can the purification kits you refer to also be used with small amounts of antibody? I ask this because your antibodies are provided in 100uL quantities.

    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/Antibody-Purification-Kit-Protein-A-ab102784.html (or use the following: https://www.abcam.com/Antibody-Purification-Kit-Protein-A-ab102784.html).

    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

    Question

    Dear Sir/Madam,
    I would like to order anti-MelanA antibody [EP1422Y] (#ab51061) and anti-MelanA antibody (#ab117440) without BSA. Is this possible?
    Thank you in advance.

    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/Antibody-Purification-kit-1-purification-ab102783.html (or use the following: https://www.abcam.com/Antibody-Purification-kit-1-purification-ab102783.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

    Question

    Thanks a lot...this really helps!

    Read More
    Answer

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

    Read More

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