Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 02, 2021

Anti-MelanA antibody [EPR20380] (ab210546)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20380] to MelanA
Suitable for: Flow Cyt, IHC-P, WB, ICC/IF
Reacts with: Mouse, Human

Product name

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

Rabbit monoclonal [EPR20380] to MelanA
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse Human
IHC-P	Mouse Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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
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.

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

pH: 7.2
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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EPR20380] (ab210546)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-MelanA antibody [EPR20380] (ab210546)

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.
Immunocytochemistry/ Immunofluorescence - Anti-MelanA antibody [EPR20380] (ab210546)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EPR20380] (ab210546)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EPR20380] (ab210546)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-MelanA antibody [EPR20380] (ab210546)

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 Cytometry - Anti-MelanA antibody [EPR20380] (ab210546)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MelanA antibody [EPR20380] (ab210546)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-MelanA antibody [EPR20380] (ab210546)

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