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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 4
#9045

Thymidylate Synthase (D5B3) XP® Rabbit mAb

Western Blotting Image 1: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
使用 Thymidylate Synthase (D5B3) XP® Rabbit mAb (上图) 或 β-Actin (13E5) Rabbit mAb #4970 (下图), 对 SK-BR-3 与 HCC827 细胞的提取位进行蛋白质印迹分析,这些细胞未经处理或经过 5-FU (1 μM) 或 Lapatinib (1 μM) 24 小时处理。
Western Blotting Image 2: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
使用 Thymidylate Synthase (D5B3) XP® Rabbit mAb (上图) 或 β-Actin (D6A8) Rabbit mAb (下图) #8457,对来自不同细胞系的提取物进行蛋白质印迹分析。
Immunohistochemistry Image 1: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
在对照肽(左图)或抗原特异性肽(右图)存在的情况下,使用 Thymidylate Synthase (D5B3) XP® Rabbit mAb 对石蜡包埋的人肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 2: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
使用 Thymidylate Synthase (D5B3) XP® Rabbit mAb 对石蜡包埋的人淋巴瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 3: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
使用 Thymidylate Synthase (D5B3) XP® Rabbit mAb 对石蜡包埋的小鼠脾脏组织进行免疫组织化学分析。
Immunohistochemistry Image 4: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
使用 Thymidylate Synthase (D5B3) XP® Rabbit mAb 对未经处理的(左图)或已经过 LY294002 #9901(右图)处理的石蜡包埋的 SK-BR-3 细胞沉淀物进行免疫组织化学分析。
Immunofluorescence Image 1: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
使用 Thymidylate Synthase (D5B3) XP® Rabbit mAb(绿色)对 LN18 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: Thymidylate Synthase (D5B3) XP® Rabbit mAb Expand Image
使用 Thymidylate Synthase (D5B3) XP® Rabbit mAb 对 HCC827 细胞(蓝色)和 HeLa 细胞(绿色)进行流式细胞分析。