WB result of PMEL/Melanoma gp100 Recombinant Rabbit mAb
Primary antibody: PMEL/Melanoma gp100 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: SK-MEL-28 whole cell lysate 20 µg
Negative control: HeLa whole cell lysate
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 70 kDa
Observed MW: 100 kDa
PMEL/Melanoma gp100 Recombinant Rabbit mAb (SDT-3393)
PMEL/Melanoma gp100 Recombinant Rabbit mAb (SDT-3393)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | PMEL/Melanoma gp100 |
| Synonyms | Melanocyte protein PMEL; ME20-M (ME20M); Melanocyte protein Pmel 17; Melanocytes lineage-specific antigen GP100; Melanoma-associated ME20 antigen; P1; D12S53E; PMEL17; SILV; PMEL |
| Location | Endosome, Melanosome, Endoplasmic reticulum membrane |
| Accession | P40967 |
| Clone Number | SDT-3393 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms |
| Positive Sample | SK-MEL-28, B16-F0 |
| Purification | Protein A |
| Concentration | 0.5 mg/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 | 1:1000-1:2000 | Hu, Ms |
| IHC-P | 1:1000 | Hu |
| ICC | 1:500 | Hu |
Background
PMEL (premelanosome protein), also called melanoma gp100, is a ~100 kDa type-I transmembrane glycoprotein encoded by the PMEL gene; it is expressed almost exclusively in pigment cells, where it is trafficked to the melanosome lumen, proteolytically processed by furin-like convertases and BACE2, and then self-assembles into non-pathological amyloid fibrils that form the striated matrix on which melanin is deposited, while concurrently its peptide fragments—particularly the gp100280–288 epitope—are presented on HLA-A*02:01 and recognized by cytotoxic T lymphocytes, making PMEL/gp100 a pivotal structural component of melanosome biogenesis and one of the most important melanoma-associated antigens targeted by immunotherapies such as tebentafusp.
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Western Blot
WB result of PMEL/Melanoma gp100 Recombinant Rabbit mAb
Primary antibody: PMEL/Melanoma gp100 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: B16-F0 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 70 kDa
Observed MW: 90 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human melanoma (case 1). Anti- PMEL/Melanoma gp100 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- PMEL/Melanoma gp100 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 colon. Anti- PMEL/Melanoma gp100 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- PMEL/Melanoma gp100 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 adenocarcinoma. Anti- PMEL/Melanoma gp100 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 cervical squamous cell carcinoma. Anti- PMEL/Melanoma gp100 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 B16-F0 cells. Anti-PMEL/Melanoma gp100 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).
