Human Thymidine Kinase 1 Antibody

Catalog #: AF8180 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF8180-100
AF8180-SP

Thymidine Kinase 1 in Human Breast Cancer Tissue.

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Human Thymidine Kinase 1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Thymidine Kinase 1 Protein in direct ELISAs.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived human Thymidine Kinase 1 protein
Ser2-Asn234
Accession # P04183

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Simple Western

20 µg/mL

A431 human epithelial carcinoma cell line and L6 rat myoblast cell line

Immunohistochemistry

1-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry

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Thymidine Kinase 1 in Human Breast Cancer Tissue. Thymidine Kinase 1 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Sheep Anti-Human Thymidine Kinase 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8180) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western

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Detection of Human and Rat Thymidine Kinase 1 by Simple Western^TM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line and L6 rat myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for Thymidine Kinase 1 at approximately 59 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human Thymidine Kinase 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8180). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Thymidine Kinase 1

Thymidine kinases are 2'-deoxythymidine kinases that phosphorylate deoxythymidine and generate deoxythymidine 5’-phosphate (1). Thymidine kinases have a key function in the synthesis of DNA and thereby in cell division, as they are part of the unique reaction chain to introduce deoxythymidine into the DNA. Two forms of thymidine kinase are present in mammalian cells, TK1 and TK2. TK1 is cell cycle-dependent and is present in the cytoplasm only in anticipation of cell division (2, 3); whereas TK2 is located in mitochondria and is cell cycle-independent (4). TK1 is synthesized by the cell during the S phase of cell division and is degraded after cell division is completed (5). TK1 normally occurs in tissue as a dimer and can be converted to more active tetrameric form by ATP binding (6). TK1 is feedback inhibited by thymidine triphosphate, the product of the further phosphorylation of thymidine (7). Because tumor cells replicate much more frequently than normal cells and requires faster DNA synthesis and higher TK1 activity, TK1 is a cancer maker especially for hematologic malignancies (8, 9). In clinical chemistry TK1 is used as a proliferation marker in the diagnosis, control of treatment and follow-up of malignant disease (10). In addition, thymidine kinase is required for the action of many antiviral drugs (11). The enzymatic activity of recombinant human TK1 is measured using a phosphatase-coupled method (12).

References

Wintersberger, E. (1997) Biochem. Soc. Trans. 25:303.
Littelfield, J.W. (1966) Biochim. Biophys. Acta 114:398.
Flemington, E, et al. (1987) Gene 52:267.
Berk, A.J. et al. (1973) Arch. Biochem. Biophys. 154:563.
Zhu, C. et al. (2006) Nucleosides Nucleotides Nucleic Acids 25:1185.
Munch-Petersen, B. et al. (1993) J. Biol. Chem. 268:15621.
Mikkelsen, N. E. et al. (2003). Biochemistry 42:5706.
Doi S, et al. (1990) Nagoya J. Med. Sci. 52:19.
Ellims PH, et al. (1981). Cancer Res. 41: 691.
Lin TS, et al. (1976) J. Med. Chem.19:495.
Baba M, et al. (1987). Biochem. Biophys. Res. Commun. 142:128.
Wu, Z.L. (2011) PLoS One 6:e23172.

Entrez Gene IDs

7083 (Human); 21877 (Mouse)

Alternate Names

EC 2.7.1.21; thymidine kinase 1 soluble isoform; Thymidine Kinase 1; thymidine kinase 1, soluble; thymidine kinase, cytosolic; thymidine kinase-1; TK1; TK2