Recombinant Anti-TRP2/DCT antibody [EPR21986] (ab221144)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21986] to TRP2/DCT
- Suitable for: mIHC, WB, IHC-P, IP
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
-
Product name
Anti-TRP2/DCT antibody [EPR21986]
See all TRP2/DCT primary antibodies -
Description
Rabbit monoclonal [EPR21986] to TRP2/DCT -
Host species
Rabbit -
Specificity
Mouse species is recommended based on WB and IP results, we do not guarantee IHC-P for mouse.
-
Tested applications
Suitable for: mIHC, WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: MeWo, SK-MEL-2, SK-MEL-28 and F9 whole cell lysates. IHC-P: Human skin and melanoma tissues. IP: SK-MEL-2 and B16-F0 whole cell lysates. mIHC: Human skin (melanocytes).
-
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
-
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, 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21986 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab221144 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 |
---|---|---|
mIHC |
1/3000.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
WB |
1/1000. Detects a band of approximately 69, 85 kDa (predicted molecular weight: 59 kDa).
|
|
IHC-P | (1) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Mouse species is recommended based on WB and IP results, we do not guarantee IHC-P for mouse. |
IP |
1/30.
|
Notes |
---|
mIHC
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 69, 85 kDa (predicted molecular weight: 59 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Mouse species is recommended based on WB and IP results, we do not guarantee IHC-P for mouse. |
IP
1/30. |
Target
-
Function
Involved in regulating eumelanin and phaeomelanin levels. -
Pathway
Pigment biosynthesis; melanin biosynthesis. -
Sequence similarities
Belongs to the tyrosinase family. -
Cellular localization
Melanosome membrane. - Information by UniProt
-
Database links
- Entrez Gene: 1638 Human
- Entrez Gene: 13190 Mouse
- Omim: 191275 Human
- SwissProt: P40126 Human
- SwissProt: P29812 Mouse
- Unigene: 301865 Human
- Unigene: 19987 Mouse
-
Alternative names
- L dopachrome tautomerase antibody
- DCT antibody
- Dopachrome delta isomerase antibody
see all
Images
-
All lanes : Anti-TRP2/DCT antibody [EPR21986] (ab221144) at 1/1000 dilution
Lane 1 : MeWo (human malignant melanoma cell line) whole cell lysate
Lane 2 : SK-MEL-2 (human skin malignant melanoma cell line) whole cell lysate
Lane 3 : SK-MEL-28 (human malignant melanoma cell line) whole cell lysate
Lane 4 : F9 (mouse embryonic testicular cancer 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
Lanes 2-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 59 kDa
Observed band size: 69,85 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time : Lane 1: 5 seconds; Lane 2: 15 seconds; Lane 3: 26 seconds; Lane 4: 3 minutes.
The molecular mass observed is consistent with what has been described in the literature (PMID: 12719423, PMID: 25009701)
-
Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling TRP2/DCT with ab221144 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in human melanoma (PMID: 24475287). 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.
-
TRP2/DCT was immunoprecipitated from 0.35 mg B16-F0 (mouse melanoma epithelial cell-like cell line) whole cell lysate with ab221144 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221144 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: B16-F0 whole cell lysate 10 µg (Input).
Lane 2: ab221144 IP in B16-F0 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab221144 in B16-F0 whole cell lysate.
Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 41 seconds.
~85KD band is the glycosylated form (PMID: 12719423)
-
Panel A: merged staining of anti-Desmoglein 1/DSG1 (gray; Opal™690), anti-Langerin (green; Opal™520) and anti-TRP2/DCT (red; Opal™570) on human skin.
Panel B: anti-TRP2/DCT stained on melanocytes.
Panel C: anti-Langerin stained on Langerhans cells.
Panel D: anti-Desmoglein 1/DSG1 stained on epidermis.The section was incubated in three rounds of staining: in the order of ab236259, ab283686, and ab221144 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
-
TRP2/DCT was immunoprecipitated from 0.35 mg SK-MEL-2 (human skin malignant melanoma cell line) whole cell lysate with ab221144 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221144 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: SK-MEL-2 whole cell lysate 10 µg (Input).
Lane 2: ab221144 IP in SK-MEL-2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab221144 in SK-MEL-2 whole cell lysate.
Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
-
Immunohistochemical analysis of paraffin-embedded human skin tissue labeling TRP2/DCT with ab221144 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in melanocytes of human skin (PMID:12535195). 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.
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
-
SDS download
-
Datasheet download
Certificate of Compliance
References (13)
ab221144 has been referenced in 13 publications.
- Ren S et al. Conjugate of ibrutinib with a TLR7 agonist suppresses melanoma progression and enhances antitumor immunity. Int J Biol Sci 18:166-179 (2022). PubMed: 34975325
- Zhao J et al. Solamargine Alleviated UVB-Induced Inflammation and Melanogenesis in Human Keratinocytes and Melanocytes via the p38 MAPK Signaling Pathway, a Promising Agent for Post-inflammatory Hyperpigmentation. Front Med (Lausanne) 9:812653 (2022). PubMed: 35770009
- Villareal MO et al. Molecular Analysis of the Melanogenesis Inhibitory Effect of Saponins-Rich Fraction of Argania spinosa Leaves Extract. Molecules 27:N/A (2022). PubMed: 36235295
- Hong C et al. Epimedium brevicornum Maxim. Extract exhibits pigmentation by melanin biosynthesis and melanosome biogenesis/transfer. Front Pharmacol 13:963160 (2022). PubMed: 36249817
- Wang X et al. Study on the Mechanism of miR-125b-5p Affecting Melanocyte Biological Behavior and Melanogenesis in Vitiligo through Regulation of MITF. Dis Markers 2022:6832680 (2022). PubMed: 36438898