WB result of MelanA Recombinant Rabbit mAb
Primary antibody: MelanA Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: OVCAR-3 whole cell lysate 20 µg
Lane 2: SK-MEL-28 whole cell lysate 20 µg
Negative control: OVCAR-3 whole cell lysate
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 13 kDa
Observed MW: 20 kDa
MelanA Recombinant Rabbit mAb (SDT-3352)
MelanA Recombinant Rabbit mAb (SDT-3352)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | MelanA |
| Synonyms | Melanoma antigen recognized by T-cells 1; MART-1; Antigen LB39-AA; Antigen SK29-AA; Protein Melan-A; MART1; MLANA |
| Location | Cytoplasm, Endoplasmic reticulum |
| Accession | Q16655 |
| Clone Number | SDT-3352 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms |
| Positive Sample | SK-MEL-28, B16F0 |
| Purification | Protein A |
| Concentration | 0.5mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000-1:5000 | Hu, Ms |
| IHC-P | 1:1000 | Hu, Ms |
| ICC | 1:500 | Hu, Ms |
Background
MelanA (also called MART-1) is a 13 kDa single-pass transmembrane protein encoded by the MLANA gene, expressed almost exclusively in melanocytes of the skin and retina, where it stabilizes and traffics the melanosomal glycoprotein gp100 to facilitate stage II melanosome biogenesis and melanin synthesis; its immunogenic 26-35 amino-acid fragment is presented on HLA class I molecules and constitutes one of the most frequent targets of circulating naïve CD8⁺ T cells, making the protein both a key histopathologic marker (detected by antibodies A103 or M2-7C10) for melanocytic tumors and a central component in experimental melanoma vaccines .
Picture
Picture
Western Blot
WB result of MelanA Recombinant Rabbit mAb
Primary antibody: MelanA Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: B16F0 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 13 kDa
Observed MW: 20 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human melanoma (case 1). Anti-MelanA antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human melanoma (case 2). Anti-MelanA antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human lung cancer. Anti-MelanA antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human ovarian cancer. Anti-MelanA antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human kidney. Anti-MelanA antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in B16F0 cells. Anti-MelanA antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ICC shows positive staining in SK-MEL-28 cells (top panel) and negative staining in OVCAR3 cells (below panel). Anti-MelanA antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
