Recombinant Anti-TRP1 antibody [EPR21960] (ab235447)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21960] to TRP1
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-TRP1 antibody [EPR21960]
See all TRP1 primary antibodies -
Description
Rabbit monoclonal [EPR21960] to TRP1 -
Host species
Rabbit -
Specificity
We recommend WB for human and mouse; we predict it will also work in WB with rat lysates but were unable to test this. -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human skin and melanoma tissues; Mouse and rat skin tissues. WB: MeWo and B16-F0 whole cell lysates. ICC/IF: MeWo and B16-F0 cells. Flow Cyt (intra): MeWo and B16-F0 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21960 -
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
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab235447 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 |
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WB |
1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 61 kDa).
We recommend WB for human and mouse; we predict it will also work in WB with rat lysates but were unable to test this. |
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IHC-P | (1) |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/1000.
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Flow Cyt (Intra) |
1/60.
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Notes |
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WB
1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 61 kDa). We recommend WB for human and mouse; we predict it will also work in WB with rat lysates but were unable to test this. |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/1000. |
Flow Cyt (Intra)
1/60. |
Target
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Function
Oxidation of 5,6-dihydroxyindole-2-carboxylic acid (DHICA) into indole-5,6-quinone-2-carboxylic acid. May regulate or influence the type of melanin synthesized. -
Tissue specificity
Pigment cells. -
Pathway
Pigment biosynthesis; melanin biosynthesis. -
Involvement in disease
Defects in TYRP1 are the cause of albinism oculocutaneous type 3 (OCA3) [MIM:203290]; also known as Rufous oculocutaneous albinism. An autosomal recessive disorder in which the biosynthesis of melanin pigment is reduced in skin, hair, and eyes. Tyrosinase activity is normal and patients have only moderate reduction of pigment. The eyes present red reflex on transillumination of the iris, dilution of color of iris, nystagmus and strabismus. Darker-skinned individuals have bright copper-red coloration of the skin and hair. -
Sequence similarities
Belongs to the tyrosinase family. -
Cellular localization
Melanosome membrane. - Information by UniProt
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Database links
- Entrez Gene: 7306 Human
- Entrez Gene: 22178 Mouse
- Entrez Gene: 298182 Rat
- Omim: 115501 Human
- SwissProt: P17643 Human
- SwissProt: P07147 Mouse
- Unigene: 270279 Human
- Unigene: 30438 Mouse
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Alternative names
- 5 antibody
- 5,6 dihydroxyindole 2 carboxylic acid oxidase antibody
- 6-dihydroxyindole-2-carboxylic acid oxidase antibody
see all
Images
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All lanes : Anti-TRP1 antibody [EPR21960] (ab235447) at 1/1000 dilution
Lane 1 : MeWo (human malignant melanoma fibroblast cell ine) whole cell lysate
Lane 2 : B16-F0 (mouse melanoma epithelial cell-like cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 61 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Exposure time: Lane 1: 10 seconds; Lane 2: 3 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The observed mass is greater than the predicted due to glycosylation.
The molecular mass observed is consistent with what has been described in the literature (PMID: 21324755; PMID: 18042623;18650849).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRP1 antibody [EPR21960] (ab235447)
Immunohistochemical analysis of paraffin-embedded human skin tissue labeling TRP1 with ab235447 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining of melanocytes in human skin (PMID:21562572). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 100% methanol fixed B16-F0 (mouse melanoma epithelial cell-like cell line) cells labeling TRP1 with ab235447 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining in the B16-F0 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized B16-F0 (mouse melanoma epithelial cell-like cell line) cells labeling TRP1 with ab235447 at 1/60 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRP1 antibody [EPR21960] (ab235447)
Immunohistochemical analysis of paraffin-embedded rat skin tissue labeling TRP1 with ab235447 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining of melanocytes in rat skin (PMID: 21562572). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRP1 antibody [EPR21960] (ab235447)
Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling TRP1 with ab235447 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining of melanocytes in mouse skin (PMID: 21562572). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRP1 antibody [EPR21960] (ab235447)
Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling TRP1 with ab235447 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in tumor cells of human melanoma (PMID: 22045183; 26068396). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MeWo (human malignant melanoma fibroblast cell line) cells labeling TRP1 with ab235447 at 1/60 (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
TRP1 is predominantly expressed in mature melanosomes (PMID: 27073483, 11266471).
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Immunofluorescent analysis of 100% methanol fixed MeWo (human malignant melanoma fibroblast cell line) cells labeling TRP1 with ab235447 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in MeWo cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (12)
ab235447 has been referenced in 12 publications.
- Wu X et al. Altered expression of ferroptosis markers and iron metabolism reveals a potential role of ferroptosis in vitiligo. Pigment Cell Melanoma Res 35:328-341 (2022). PubMed: 35218147
- Yang CY et al. UVB-Induced Secretion of IL-1β Promotes Melanogenesis by Upregulating TYR/TRP-1 Expression In Vitro. Biomed Res Int 2022:8230646 (2022). PubMed: 35572734
- Tian H et al. Induced retinal pigment epithelial cells with anti-epithelial-to-mesenchymal transition ability delay retinal degeneration. iScience 25:105050 (2022). PubMed: 36185374
- Hong C et al. Epimedium brevicornum Maxim. Extract exhibits pigmentation by melanin biosynthesis and melanosome biogenesis/transfer. Front Pharmacol 13:963160 (2022). PubMed: 36249817
- Zhang Y et al. Establishment and validation of evaluation models for post-inflammatory pigmentation abnormalities. Front Immunol 13:991594 (2022). PubMed: 36389813