Anti-Melanoma gp100 antibody [HMB45] (ab787)
- Datasheet
- References (13)
- Protocols
Overview
-
Product nameAnti-Melanoma gp100 antibody [HMB45]
See all Melanoma gp100 primary antibodies -
DescriptionMouse monoclonal [HMB45] to Melanoma gp100
-
Host speciesMouse
-
Tested applicationsSuitable for: Flow Cyt, IHC-P, ICC/IFmore details
-
Species reactivityReacts with: Human
Does not react with: Rat, Dog -
Immunogen
Tissue, cells or virus corresponding to Human Melanoma. BALB/C mice were injected with extract of pigmented malanoma metastases from lymph nodes.
Database link: P40967 -
Positive control
- IHC-P: Human melanoma and testis tissue.
Properties
-
FormLiquid
-
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
-
Storage bufferPreservative: 0.05% Sodium azide
Constituents: Tissue culture supernatant, 0.05% BSA -
PurityTissue culture supernatant
-
ClonalityMonoclonal
-
Clone numberHMB45
-
IsotypeIgG1
-
Light chain typekappa
-
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
Applications
Our Abpromise guarantee covers the use of ab787 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 |
|---|---|---|
| Flow Cyt | 1/10. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
|
| IHC-P | Use a concentration of 0.5 - 1 µg/ml. Incubate for 30 minutes at room temperature. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 minutes |
|
| ICC/IF | Use a concentration of 0.5 - 1 µg/ml. PubMed: 19841138 |
Target
-
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. -
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 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
-
Database links
- Entrez Gene: 6490 Human
- Omim: 155550 Human
- SwissProt: P40967 Human
- Unigene: 95972 Human
-
Alternative names
- 95 kDa melanocyte specific secreted glycoprotein antibody
- 95 kDa melanocyte-specific secreted glycoprotein antibody
- D12S53E antibody
see all
Images
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)](https://www.abcam.com/ps/products/0/ab787/Images/ab787-302927-787a.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)Formalin-fixed, paraffin-embedded human melanoma tissue stained with ab787 at 1 μg/ml in immunohistochemical analysis.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)](https://www.abcam.com/ps/products/0/ab787/Images/ab787_1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)ab787 - immunohistochemistry
Formalin fixed paraffin embedded human melanoma stained with ab787, using ABC and AEC chromogen.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)](https://www.abcam.com/ps/products/0/ab787/Images/ab787-302928-787b.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)Formalin-fixed, paraffin-embedded human testis tissue stained with ab787 at 1 μg/ml in immunohistochemical analysis.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)](https://www.abcam.com/ps/products/0/ab787/Images/Melanoma-Primary-antibodies-ab787-2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)This image is courtesy of an Abreview submitted by Dr. Hongwei Shaoab787 staining Melanoma in the human Melanoma cell line WM3248 xenopraph by IHC-P (formaldehyde-fixed paraffin-embedded sections). Tissue samples were fixed with formaldehyde; permeabilized with 0.1% Triton X-100 and blocked with 100% Dakocytomation X0909 for 1 hour at room temperature; antigen retrival was by heat mediation in Citric buffer (pH6). The sample was incubated with primary antibody (1/25) at 4°C for 18 hours. An HRP-conjugated Goat polyclonal to mouse IgG (1/100) was used as secondary antibody. -
![Flow Cytometry - Anti-Melanoma gp100 antibody [HMB45] (ab787)](https://www.abcam.com/ps/products/0/ab787/Images/Melanoma-Primary-antibodies-ab787-3.jpg)
Flow Cytometry - Anti-Melanoma gp100 antibody [HMB45] (ab787)Overlay histogram showing Malme-3 cells stained with ab787 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab787, 1/10 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Malme-3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
Protocols
References
This product has been referenced in:
- Bergam P et al. ABCB6 Resides in Melanosomes and Regulates Early Steps of Melanogenesis Required for PMEL Amyloid Matrix Formation. J Mol Biol N/A:N/A (2018). Read more (PubMed: 29940187) »
- Sweeny L et al. Antiangiogenic antibody improves melanoma detection by fluorescently labeled therapeutic antibodies. Laryngoscope 126:E387-E395 (2016). Read more (PubMed: 27576611) »
