Recombinant Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
- Datasheet
- References (6)
- Protocols
Overview
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Product name
Anti-Melanoma gp100 antibody [EPR4864]
See all Melanoma gp100 primary antibodies -
Description
Rabbit monoclonal [EPR4864] to Melanoma gp100 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-Fr, WB, IHC-Pmore details
Unsuitable for: ICC or IP -
Species reactivity
Reacts with: Human -
Immunogen
within Human Melanoma gp100 aa 600 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P40967 -
Positive control
- Human melanoma tissue lysate; Human melanoma tissue ;Placenta tissue; Normal skin tissue;Normal kidney tissue.
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
This product is a recombinant rabbit monoclonal antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20ºC. -
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
EPR4864 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab137062 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 |
|---|---|---|
| IHC-Fr | Use at an assay dependent concentration. | |
| WB | 1/1000 - 1/10000. Detects a band of approximately 100, 26 kDa (predicted molecular weight: 70 kDa). For unpurified, use 1/1000 - 1/2000. |
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| IHC-P | 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. For unpurified, use 1/50 - 1/100. |
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
- Omim: 155550 Human
- SwissProt: P40967 Human
- Unigene: 95972 Human
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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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![Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-239818-anti-melanoma-gp100-antibody-epr4864-western-blot.jpg)
Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)Anti-Melanoma gp100 antibody [EPR4864] (ab137062) at 1/10000 dilution (purified) + Human melanoma tissue lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 70 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Additional bands at: 26 kDa. We are unsure as to the identity of these extra bands.Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-239820-anti-melanoma-gp100-antibody-epr4864-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)Immunohistochemical staining of paraffin embedded human melanoma with purified ab137062 at a working dilution of 1 in 400. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
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![Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-151580-anti-melanoma-gp100-antibody-epr4864-western-blot.jpg)
Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)Anti-Melanoma gp100 antibody [EPR4864] (ab137062) at 1/1000 dilution (unpurified) + Human melanoma tissue lysate at 10 µg
Secondary
HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution
Predicted band size: 70 kDa -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-239819-anti-melanoma-gp100-antibody-epr4864-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)Immunohistochemical staining of paraffin embedded human melanoma with unpurified ab137062 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
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![Immunohistochemistry (Frozen sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-226356-anti-melanoma-gp100-antibody-epr4864-immunohistochemistry.jpg)
Immunohistochemistry (Frozen sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)This image is courtesy of an anonymous AbreviewUnpurified ab137062 staining melanoma gp100 in human melanoma tissue sections by immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized and blocked with 4% horse serum + 1% BSA for 30 minutes at 22°C. Samples were incubated with primary antibody (1/500 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 568-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Magnification: 20X. Counterstained with DAPI.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-151585-anti-melanoma-gp100-antibody-epr4864-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)Immunohistochemical analysis using unpurified ab137062 showing negative staining in placenta tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-151584-anti-melanoma-gp100-antibody-epr4864-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)Immunohistochemical analysis using unpurifed ab137062 showing negative staining in normal skin tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-151583-anti-melanoma-gp100-antibody-epr4864-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)Immunohistochemical analysis using unpurified ab137062 showing negative staining in normal kidney tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)](https://www.abcam.com/ps/products/137/ab137062/Images/ab137062-151581-anti-melanoma-gp100-antibody-epr4864-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)Immunohistochemical analysis using unpurified ab137062 showing negative staining in normal stomach tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
References
This product has been referenced in:
- Graham M et al. Repeat domain-associated O-glycans govern PMEL fibrillar sheet architecture. Sci Rep 9:6101 (2019). Read more (PubMed: 30988362) »
- Yi WJ et al. Degraded melanocores are incompetent to protect epidermal keratinocytes against UV damage. Cell Cycle 17:844-857 (2018). Read more (PubMed: 29623762) »
