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: WB, IHC-P, ICC/IF, Flow Cyt
- Reacts with: Human
Product nameAnti-MelanA antibody [EP1422Y]
See all MelanA primary antibodies
DescriptionRabbit monoclonal [EP1422Y] to MelanA
Tested applicationsSuitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
Species reactivityReacts with: Human
Synthetic peptide within Human MelanA aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: Q16655
- WB: Human melanoma lysates. IHC-P: Human melanoma tissue. Flow Cyt: A375 and Malme-3M cells. ICC/IF: MeWo cells
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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
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.
Storage instructionsShipped 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.
Dissociation constant (KD)KD = 1.27 x 10 -9 M Learn more about KD
Storage bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|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.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionInvolved 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 specificityExpression is restricted to melanoma and melanocyte cell lines and retina.
Cellular localizationEndoplasmic 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
- Antigen LB39 AA antibody
- Antigen LB39-AA antibody
- Antigen SK29 AA antibody
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 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.
Anti-MelanA antibody [EP1422Y] (ab51061) at 1/20000 dilution (Purified) + Human melanoma lysates at 15 µg
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 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).
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.
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KD
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.
ab51061 has been referenced in 21 publications.
- Rosenbaum SR et al. FOXD3 Regulates VISTA Expression in Melanoma. Cell Rep 30:510-524.e6 (2020). PubMed: 31940493
- Sprouse ML et al. PMN-MDSCs Enhance CTC Metastatic Properties through Reciprocal Interactions via ROS/Notch/Nodal Signaling. Int J Mol Sci 20:N/A (2019). PubMed: 31003475
- Regazzetti C et al. Melanocytes Sense Blue Light and Regulate Pigmentation through Opsin-3. J Invest Dermatol 138:171-178 (2018). PubMed: 28842328
- Vishnoi M et al. Targeting USP7 Identifies a Metastasis-Competent State within Bone Marrow-Resident Melanoma CTCs. Cancer Res 78:5349-5362 (2018). PubMed: 30026332
- Wang L et al. Aberrant SIRT6 expression contributes to melanoma growth: Role of the autophagy paradox and IGF-AKT signaling. Autophagy 14:518-533 (2018). PubMed: 29215322