Product nameAnti-Melanoma gp100 antibody [EP4863(2)] (HRP)
See all Melanoma gp100 primary antibodies
DescriptionRabbit monoclonal [EP4863(2)] to Melanoma gp100 (HRP)
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Mouse, Human
Synthetic peptide within Human Melanoma gp100 aa 1-100. The exact sequence is proprietary.
Database link: P40967
- WB: B16-F0 cell lysate, human melanoma tissue lysate. IHC-P: FFPE Human skin melanoma.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. Store In the Dark.
Storage bufferpH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: 30% Glycerol, 1% BSA, PBS
Concentration information loading...
- Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)
- Anti-Melanoma gp100 antibody [EP4863(2)] - Low endotoxin, Azide free (ab193201)
- Anti-Melanoma gp100 antibody [EP4863(2)] (Biotin) (ab204674)
- Anti-Melanoma gp100 antibody [EP4863(2)] (Alexa Fluor® 647) (ab246730)
- Anti-Melanoma gp100 antibody [EP4863(2)] (PE) (ab246731)
- Anti-Melanoma gp100 antibody [EP4863(2)] - BSA and Azide free (ab256497)
Our Abpromise guarantee covers the use of ab205469 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 70 kDa).|
FunctionPlays a central role in the biogenesis of melanosomes. Involved in the maturation of melanosomes from stage I to II. The transition from stage I melanosomes to stage II melanosomes involves an elongation of the vesicle, and the appearance within of distinct fibrillar structures. Release of the soluble form, ME20-S, could protect tumor cells from antibody mediated immunity.
Tissue specificityPreferentially expressed in melanomas. Some expression was found in dysplastic nevi. Not found in normal tissues nor in carcinomas. Normally expressed at low levels in quiescent adult melanocytes but overexpressed by proliferating neonatal melanocytes and during tumor growth.
Sequence similaritiesBelongs to the PMEL/NMB family.
Contains 1 PKD domain.
DomainThe RPT domain is essential for the generation of the fibrillar matrix of melanosomes.
The lumenal domain is necessary for correct processing and trafficking to melanosomes.
modificationsA small amount of P1/P100 (major form) undergoes glycosylation to yield P2/P120 (minor form). P2 is cleaved by a furin-like proprotein convertase (PC) in a pH-dependent manner in a post-Golgi, prelysosomal compartment into two disulfide-linked subunits: a large lumenal subunit, M-alpha/ME20-S, and an integral membrane subunit, M-beta. Despite cleavage, only a small fraction of M-alpha is secreted, whereas most M-alpha and M-beta remain associated with each other intracellularly. M-alpha is further processed to M-alpha N and M-alpha C. M-alpha C further undergoes processing to yield M-alpha C1 and M-alpha C3 (M-alpha C2 in the case of PMEL17-is or PMEL17-ls). Formation of intralumenal fibrils in the melanosomes requires the formation of M-alpha that becomes incorporated into the fibrils. Stage II melanosomes harbor only Golgi-modified Pmel17 fragments that are derived from M-alpha and that bear sialylated O-linked oligosaccharides.
N-glycosylated. O-glycosylated; contains sialic acid.
Cellular localizationSecreted and Endoplasmic reticulum membrane. Golgi apparatus. Melanosome. Endosome > multivesicular body. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Localizes predominantly to intralumenal vesicles (ILVs) within multivesicular bodies. Associates with ILVs found within the lumen of premelanosomes and melanosomes and particularly in compartments that serve as precursors to the striated stage II premelanosomes.
- Information by UniProt
- 95 kDa melanocyte specific secreted glycoprotein antibody
- 95 kDa melanocyte-specific secreted glycoprotein antibody
- D12S53E antibody
All lanes : Anti-Melanoma gp100 antibody [EP4863(2)] (HRP) (ab205469) at 1/1000 dilution
Lane 1 : B16-F0 (Mouse) Whole Cell Lysate
Lane 2 : Melanoma (Human) Whole Cell Lysate - tumor tissue
Lysates/proteins at 10 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutes
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab205469 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
IHC image of gp100 staining in a section of formalin-fixed paraffin-embedded Human Skin Melanoma*, performed on a Leica BONDTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab205469, 1/50 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
ab205469 has not yet been referenced specifically in any publications.