TRP2 / DCT Recombinant Rabbit Monoclonal Antibody [PSH25-05]
cat.: HA724499
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IHC-P, FC(Intra), IP |
| Clonality: | Monoclonal |
| Clone number: | PSH25-05 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 59 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human TRP2 / DCT aa 1-250. |
| Positive control: | MeWo (Human melanoma cell) cell lysate, SK-MEL-28 (Human malignant melanoma cell) cell lysate, B16-F1 (Mouse melanoma cell) cell lysate, Mouse skin tissue lysate. |
| Subcellular location: | Melanosome membrane, Melanosome. |
| Recommended Dilutions:
WB IHC-P FC(Intra) IP |
1:20,000 1:50-1:200 1:5,000 1-2μg/sample |
| Uniprot #: | SwissProt: P40126 Human | P29812 Mouse |
| Alternative names: | DCT TYRP2 L-dopachrome tautomerase DCT DT L-dopachrome Delta-isomerase Tyrosinase-related protein 2 TRP-2 TRP2 |
Images
|
Fig1:
Western blot analysis of TRP2 / DCT on different lysates with Rabbit anti-TRP2 / DCT antibody (HA724499) at 1/20,000 dilution.
Lane 1: MeWo (Human melanoma cell) cell lysate (15 µg/Lane) Lane 2: SK-MEL-28 (Human malignant melanoma cell) cell lysate (15 µg/Lane) Lane 3: U-2 OS (Human osteosarcoma cell) cell lysate (15 µg/Lane) Lane 4: B16-F1 (Mouse melanoma cell) cell lysate (15 µg/Lane) Lane 5: Mouse skin tissue lysate (30 µg/Lane) Exposure time: 12 seconds; ECL: K1801 Negative expression of TRP2 / DCT protein in U-2 OS cell is consistent with the predicted expression pattern Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA724499, 1/20,000 in primary antibody dilution buffer (K1803), overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 59 kDa Observed band size: 69, 85 kDa |
|
Fig2:
Application: Immunohistochemistry (IHC-P)
Species: Human Tissue: Melanoma Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA724499, 1/50 (left), TBST(-) (right), 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig3:
Application: Immunohistochemistry (IHC-P)
Species: Human Tissue: Skin Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA724499, 1/50, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig4:
Application: Immunohistochemistry (IHC-P)
Species: Mouse Tissue: Skin Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA724499, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig5:
Immunoprecipitation (IP)
TRP2 / DCT was immunoprecipitated in 0.2 mg SK-MEL-28 (Human malignant melanoma cell) cell lysate with HA724499 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724499 at 1/10,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SK-MEL-28 cell lysate (input) Lane 2: HA724499 IP in SK-MEL-28 cell lysate Lane 3: Rabbit IgG instead of HA724499 in SK-MEL-28 cell lysate Exposure time: 4 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA724499, 1/10,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 59 kDa Observed band size: 69, 85 kDa |
|
Fig6:
Application: Flow Cytometry (Intra)
Species: Human Sample: U-2 OS (Human osteosarcoma cell)/SK-MEL-28 (Human malignant melanoma cell) Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. Permeabilization: 0.1% Tween-20, 15 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. Antibody dilution buffer: 1x PBS. Primary antibody: HA724499(1/5,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature. Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature. Negative expression of TRP2 / DCT protein in U-2 OS cell is consistent with the predicted expression pattern |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.