Recombinant Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)
![Immunoprecipitation - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)](https://www.abcam.com/ps/products/137/ab137078/Images/ab137078-414315-AntiMelanoma-gp100-antibody.jpg "Immunoprecipitation - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP4863(2)] to Melanoma gp100
- Suitable for: WB, IHC-P, IP, Flow Cyt (Intra), IA, ICC/IF
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Melanoma gp100 antibody [EP4863(2)]
See all Melanoma gp100 primary antibodies -
Description
Rabbit monoclonal [EP4863(2)] to Melanoma gp100 -
Host species
Rabbit -
Specificity
The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
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Tested applications
Suitable for: WB, IHC-P, IP, Flow Cyt (Intra), IA, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide within Human Melanoma gp100 aa 1-100. The exact sequence is proprietary.
Database link: P40967 -
Positive control
- WB: B16-F0 (Mouse melanoma epithelial cell-like) whole cell lysate and Human melanoma lysate. IHC: Human melanoma tissue. ICC/IF: B16-F0 (Mouse melanoma epithelial cell-like) cell line. Flow Cyt (intra): B16-F0 (Mouse melanoma epithelial cell-like) cell line. IP: B16-F0 (Mouse melanoma epithelial cell-like) whole cell lysate.
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General notes
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP4863(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-Melanoma gp100 antibody [EP4863(2)] - Low endotoxin, Azide free (ab193201)
- Biotin Anti-Melanoma gp100 antibody [EP4863(2)] (ab204674)
- HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469)
- Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)] (ab246730)
- PE Anti-Melanoma gp100 antibody [EP4863(2)] (ab246731)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab137078 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 70 kDa.
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| IHC-P | (5) |
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse. |
| IP |
1/30.
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| Flow Cyt (Intra) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IA |
1/50.
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| ICC/IF | (2) |
1/50.
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| Notes |
|---|
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WB
1/1000 - 1/10000. Predicted molecular weight: 70 kDa. |
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IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse. |
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IP
1/30. |
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Flow Cyt (Intra)
1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IA
1/50. |
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ICC/IF
1/50. |
Target
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Function
Plays 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 specificity
Preferentially 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 similarities
Belongs to the PMEL/NMB family.
Contains 1 PKD domain. -
Domain
The 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. -
Post-translational
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 localization
Secreted 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
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Database links
- Entrez Gene: 6490 Human
- Entrez Gene: 20431 Mouse
- Omim: 155550 Human
- SwissProt: P40967 Human
- SwissProt: Q60696 Mouse
- Unigene: 95972 Human
- Unigene: 16756 Mouse
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Alternative names
- 95 kDa melanocyte specific secreted glycoprotein antibody
- 95 kDa melanocyte-specific secreted glycoprotein antibody
- D12S53E antibody
see all
Images
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Immunoprecipitation - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)
ab137078 immunoprecipitating Melanoma gp100. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.
Lane 1: B16-F0 (mouse melanoma epithelial cell-like) whole cell lysate 10ug
Lane 2: B16-F0 (mouse melanoma epithelial cell-like) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab137078 in B16-F0 whole cell lysateExposure time: 1 second
Observed MW: 85-100 KDa
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![Flow Cytometry (Intracellular) - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)](https://www.abcam.com/ps/products/137/ab137078/Images/ab137078-414314-AntiMelanoma-gp100-antibody.jpg)
Flow Cytometry (Intracellular) - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)Overlay histogram showing B16-F0 (Mouse skin melanoma cell) cells stained with ab137078 (red line). The cells were fixed with 4% paraformaldehydeand then permeabilized with 90% methanol. The cells were then incubated with ab137078 at 1/40 dilution. The secondary antibody used wasGoat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution. Isotype control antibody (black line) was Rabbit monoclonal IgG (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody.
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![Immunocytochemistry/ Immunofluorescence - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)](https://www.abcam.com/ps/products/137/ab137078/Images/ab137078-414313-AntiMelanoma-gp100-antibody.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)ab137078 staining Melanoma gp100 in B16-F0 (mouse skin) cells by ICC (Immunocytochemistry). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/50. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) ab195889 was used as a counterstain at a dilution of 1/200. DAPI was used as a nuclear counterstain.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)](https://www.abcam.com/ps/products/137/ab137078/Images/ab137078-414312-AntiMelanoma-gp100-antibody.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)ab137078 staining Melanoma gp100 in human melanoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250 for 30 minutes at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody.
The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.
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![Western blot - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)](https://www.abcam.com/ps/products/137/ab137078/Images/ab137078-414311-AntiMelanoma-gp100-antibody.jpg)
Western blot - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078) at 1/1000 dilution + Human melanoma lysate 15µg
Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 70 kDaExposure time: 18-0 seconds
Observed MW: 85-100 KDa
The molecular weights observed are consistent with what have been described in PMID: 16492709 and PMID: 7961886
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![Western blot - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)](https://www.abcam.com/ps/products/137/ab137078/Images/ab137078-414310-AntiMelanoma-gp100-antibody.jpg)
Western blot - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078) at 1/10000 dilution + B16-F0 (Mouse melanoma epithelial cell-like) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 70 kDaExposure time: 50 seconds
Observed MW: 85-100 KDa
The molecular weights observed are consistent with what have been described in PMID: 16492709 and PMID: 7961886
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![Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)](https://www.abcam.com/ps/products/137/ab137078/Images/ab137078-18-benefits-of-recombinant-antibodies.png)
Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (31)
ab137078 has been referenced in 31 publications.
- Ostojic J et al. Transcriptional co-activator regulates melanocyte differentiation and oncogenesis by integrating cAMP and MAPK/ERK pathways. Cell Rep 35:109136 (2021). PubMed: 34010639
- Rosenbaum SR et al. FOXD3 Regulates VISTA Expression in Melanoma. Cell Rep 30:510-524.e6 (2020). PubMed: 31940493
- Sun L et al. Syntenin regulates melanogenesis via the p38 MAPK pathway. Mol Med Rep 22:733-738 (2020). PubMed: 32626944
- Zurina IM et al. Human Melanocyte-Derived Spheroids: A Precise Test System for Drug Screening and a Multicellular Unit for Tissue Engineering. Front Bioeng Biotechnol 8:540 (2020). PubMed: 32582665
- Ruiz-Vega R et al. Dynamics of nevus development implicate cell cooperation in the growth arrest of transformed melanocytes. Elife 9:N/A (2020). PubMed: 33047672