Revision 4
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

30

Source/Isotype:

Rabbit 

UniProt ID:

#P04818

Entrez-Gene Id:

7298

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:200
Immunofluorescence (Immunocytochemistry) 1:50
Flow Cytometry (Fixed/Permeabilized) 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #81327.

Specificity / Sensitivity

Thymidylate Synthase (D5B3) XP® Rabbit mAb detects endogenous levels of total thymidylate synthase protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human thymidylate synthase protein.

Background

The methylation of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP) is an essential step in the formation of thymine nucleotides (1,2, reviewed in 3). This process is catalyzed by thymidylate synthase (TS or TYMS), a homodimer composed of two 30 kDa subunits. TS is an intracellular enzyme that provides the sole de novo source of thymidylate, making it a required enzyme in DNA biosynthesis with activity highest in proliferating cells (1). Being the exclusive source of dTMP, investigators have concluded that TS is also an important target for anticancer agents such as 5-fluorouracil (5-FU) (1-5). 5-FU acts as a TS inhibitor and is active against solid tumors such as colon, breast, head, and neck. Research studies have demonstrated that patients with metastases expressing lower levels of TS have a higher response rate to treatment with 5-FU than patients with tumors that have increased levels of TS (5). Researchers continue to investigate TS expression in different types of cancers (6-10).
Levels of thymidylate synthase can be modultated in vitro by treatment with various drugs, such as 5-FU and lapatinib. 5-FU treatment induces the formation of two distinct bands, one at 35 kDa and another at 38 kDa, while lapatinib treatment results in a decrease in thymidylate synthase expression (11,12).

  1. Johnston, P.G. et al. (1991) Cancer Res 51, 6668-76.
  2. Aschele, C. et al. (2002) Ann Oncol 13, 1882-92.
  3. Jackman, A.L. and Calvert, A.H. (1995) Ann Oncol 6, 871-81.
  4. Van Triest, B. et al. (2000) J Histochem Cytochem 48, 755-60.
  5. Johnston, P.G. et al. (1994) J Clin Oncol 12, 2640-7.
  6. Kwon, H.C. et al. (2007) Ann Oncol 18, 504-9.
  7. Allegra, C.J. et al. (2002) J Clin Oncol 20, 1735-43.
  8. Allegra, C.J. et al. (2003) J Clin Oncol 21, 241-50.
  9. Tsourouflis, G. et al. (2008) Dig Dis Sci 53, 1289-96.
  10. Kim, S.H. et al. (2009) Am J Clin Oncol 32, 38-43.
  11. Chu, E. et al. (1993) Mol Pharmacol 43, 527-33.
  12. Kim, H.P. et al. (2009) PLoS One 4, e5933.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 4
#9045

Thymidylate Synthase (D5B3) XP® Rabbit mAb

Western Blotting Image 1: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from SK-BR-3 and HCC827 cells, untreated or treated with 5-FU (1 μM) or lapatinib (1 μM) for 24 hr, using Thymidylate Synthase (D5B3) XP® Rabbit mAb (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower).
Western Blotting Image 2: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using Thymidylate Synthase (D5B3) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb (lower) #8457.
Immunohistochemistry Image 1: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Thymidylate Synthase (D5B3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemistry Image 2: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lymphoma using Thymidylate Synthase (D5B3) XP® Rabbit mAb.
Immunohistochemistry Image 3: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using Thymidylate Synthase (D5B3) XP® Rabbit mAb.
Immunohistochemistry Image 4: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded SK-BR-3 cell pellets, untreated (left) or treated with LY294002 #9901 (right), using Thymidylate Synthase (D5B3) XP® Rabbit mAb.
Immunofluorescence Image 1: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of LN18 cells using Thymidylate Synthase (D5B3) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
Flow cytometric analysis of HCC827 cells (blue) and HeLa cells (green) using Thymidylate Synthase (D5B3) XP® Rabbit mAb.