td-Tomato
| Cat Number: | MAB-94632 |
|---|---|
| Conjugate: | Unconjugated |
| Size: | 100 ug |
| Clone: | T5A6 |
| Concentration: | 1mg/mL |
| Host: | Ms |
| Isotype: | IgG1 |
| Immunogen: | length recombinant td Tomato protein expressed in and purified from E. coli |
| Reactivity: | All Species |
| Applications: | Western blot: 1:1,000 |
| Molecular Weight: | 28kDa |
| Purification: | Aff. Pur. |
| Background: | The td-Tomato protein is derived from DsRed, a red fluorescent protein from so-called disc corals of the genus Discosoma. DsRed is similar in size and properties to GFP, but, obviously, produces a red rather than a green fluorochrome. The original DsRed was engineered extensively in the Tsien lab to prevent it from forming tetramers and dimers and to modify and improve the spectral properties (1-3). Several further cycles of mutation, directed modification and evolutionary selection produced td-Tomato , which has an excitation maximum at 587 nm and and emission maximum at 610 nm (4). |
| Form: | Liquid |
| Buffer: | Purified at 1mg/mL in 50% PBS, 50% glycerol, 5mM NaN3 |
| Storage: | Shipped on ice. Store at 4°C. For long term storage, leave frozen at -20°C. Avoid freeze / thaw cycles. |
Western blot analysis of various HEK293 cell lysates using mouse
mAb to td-Tomato dilution 1:5,000, in green. [1] protein
standard, [2] untransfected HEK293 lysate, [3] lysate of HEK293
cells transfected with td-Tomato-HA construct, and [4] lysate of
HEK293 cells transfected with eGFP construct. As expected from our
epitope mapping data, T5A6 does not recognize eGFP. The major
band at about 28kDa corresponds to the full length td-Tomato protein
and the lower band at about 21kDa is an td-Tomato breakdown
product.
Immunofluorescent analysis of HEK293 cells stably transduced with
a lentiviral vector expressing td-Tomato-HA construct (red) and
stained with mouse mAb to td-Tomato, T5A6, dilution 1:500 in
green. The blue is Hoechst staining of nuclear DNA. The T5A6
antibody reveals the td-Tomato protein expressed only in transduced
cells which appear golden in color. Untransduced cells expressing no
td-Tomato are not recognized by the T5A6 antibody and so only
their nuclei are visible.
References
1. Matz MV, et al. Fluorescent proteins fromnonbioluminescent Anthozoa species. Nat.Biotechnol. 17:969-73 (1999)
2. Baird GS, Zacharias DA, Tsien RY.Biochemistry, mutagenesis, and oligomerizationof DsRed, a red fluorescent protein from coral.
PNAS 97:11984-9 (2000).
3. Chalfie M, et al. Green fluorescent protein asa marker for gene expression. Science263:802-5 (1994).
4. Gross LA. et al. The structure of thechromophore within DsRed, a red fluorescentprotein from coral. PNAS 97:11990-5 (2000).
5. Heikal AA. et al. Molecular spectroscopy anddynamics of intrinsically fluorescent proteins:coral red (dsRed) and yellow (Citrine). PNAS97:11996-2001 (2000).
6. Shaner NC. et al. Improved monomeric red,orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein.Nat. Biotech. 22:1567-72 (2004