Recombinant
RabMAb

Recombinant Anti-MelanA antibody [EPR20380] (ab210546)

Rabbit recombinant monoclonal MelanA antibody [EPR20380]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Human. Independently reviewed in 4 review(s).

Overview

  • Product name

    Anti-MelanA antibody [EPR20380]
    See all MelanA primary antibodies
  • Description

    Rabbit monoclonal [EPR20380] to MelanA
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, IHC-P, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human MelanA aa 50 to the C-terminus. The exact sequence is proprietary.
    Database link: Q16655

  • Positive control

    • WB: Human melanoma lysate; B16F0 and MeWo whole cell lysates. IHC-P: Human malignant melanoma and metastatic malignant melanoma tissues; Mouse hair bulb tissue. ICC/IF: MeWo and B16F0 cells. Flow Cyt: B16F0 cells.
  • General notes

    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

  • Form

    Liquid
  • Storage instructions

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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20380
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab210546 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 1/500.

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

 

IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 18 kDa (predicted molecular weight: 13 kDa).
ICC/IF 1/1000.

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

  • Immunohistochemical analysis of paraffin-embedded human malignant melanoma tissue labeling MelanA with ab210546 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on human malignant melanoma is observed [PMID: 17445277] [PMID: 21840568].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MeWo (Human malignant melanoma cell line) cells labeling MelanA with ab210546 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on MeWo cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized B16F0 (Mouse melanoma cell line) cells labeling MelanA with ab210546 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on B16F0 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human metastatic malignant melanoma tissue labeling MelanA with ab210546 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on human metastatic malignant melanoma is observed [PMID: 17445277] [PMID: 21840568].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse hair bulb tissue labeling MelanA with ab210546 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on melanocytes of mouse hair bulb is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • All lanes : Anti-MelanA antibody [EPR20380] (ab210546) at 1/2000 dilution

    Lane 1 : Human melanoma lysate
    Lane 2 : A-375 (Human malignant melanoma cell line) whole cell lysate
    Lane 3 : B16F0 (Mouse melanoma cell line) whole cell lysate
    Lane 4 : MeWo (Human malignant melanoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 13 kDa
    Observed band size: 18 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 10 seconds; Lane 3 and 4: 3 seconds.

    A-375 doesn’t express MelanA. The expression profile observed is consistent with what has been described in the literature (PMID: 12670905).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed B16F0 (Mouse melanoma cell line) cells labeling MelanA with ab210546 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling MelanA with ab210546 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Negative control: no staining on human liver cancer [PMID: 10096358].

    Counter stained with Hematoxylin.

References

This product has been referenced in:

  • Chung HJ  et al. Pattern analysis of 532- and 1064-nm microlens array-type, picosecond-domain laser-induced tissue reactions in ex vivo human skin. Lasers Med Sci N/A:N/A (2019). Read more (PubMed: 30604347) »
See 1 Publication for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (melanoma xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
melanoma xenograft
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF

Abcam user community

Verified customer

Submitted Aug 06 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (melanoma xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
melanoma xenograft
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF

Abcam user community

Verified customer

Submitted Aug 06 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (skin)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
skin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Miss. Jin Young Park

Verified customer

Submitted Nov 12 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
skin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Miss. Jin Young Park

Verified customer

Submitted Aug 31 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (BRaf(V600E) Pten(del) Tyr-CreERT2 cutaneous melano)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9, Tris EDTA
Permeabilization
No
Specification
BRaf(V600E) Pten(del) Tyr-CreERT2 cutaneous melano
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jun 06 2018

Application
ELISA
Sample
Human Recombinant protein (melanoma patient serum)
Specification
melanoma patient serum
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Type
Direct

Frau. Mirjam Fässler

Verified customer

Submitted Sep 20 2017

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