Recombinant Human TSH alpha/beta Heterodimer Protein

• Reactivity: Hu
• Applications: Bioactivity

Recombinant Human TSH alpha/beta Heterodimer Protein Summary

• Details of Functionality: Measured by its ability to induce cAMP accumulation in HEK293 human embryonic kidney cells transfected with human TSH R. Morgenthaler, N.G. et al. (1998) Horm. Metab. Res. 30:162. The ED₅₀ for this effect is 0.25-1.5 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human TSH alpha/beta Heterodimer protein
  

  Human TSH alpha (Ala25-Ser116) Accession # P01215

  Human TSH beta (Phe21-Val138) Accession # P01222
  N-terminus	C-terminus

• Accession #: P01215 (TSH alpha ) & P01222 (TSH beta )
• N-terminal Sequence: Ala25 (TSH alpha ) & Phe21 (TSH beta )
• Structure / Form: Noncovalently-linked heterodimer
• Protein/Peptide Type: Recombinant Proteins
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 10.2 kDa (TSH alpha ), 13.5 kDa (TSH beta ).
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 15-29 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human TSH alpha/beta Heterodimer Protein

• thyroid stimulating hormone beta
• Thyrotropin
• TSH alpha/beta Heterodimer
• TSHB
• TSH-B
• TSH-BETA

Background

TSH (thyroid stimulating hormone), also known as thyrotropin, is a member of the cysteine knot growth factor superfamily (1-4). It is a heterodimer of a 15 kDa unique subunit, TSH beta , with a 14 kDa alpha subunit, CGa (common glycoprotein hormone alpha) that is shared with lutropin (LH), follitropin (FSH) and chorionic gonadotropin (CG) (1-4). Beta subunits of the four glycoprotein hormones share 37-43% amino acid (aa) identity. Mature human TSH  beta shares 92%, 90%, 90%, 89%, 89%, 89%, and 88% aa identity with canine, rat, equine, mouse, bovine, porcine, and feline TSH beta , respectively. Mature human CGa shares 69%-73% aa identity with canine, rabbit, rat, mouse, bovine, ovine, porcine, feline and equine CGa. Each subunit forms a cysteine knot structure with three disulfide bridges (1). A loop of the TSH beta subunit, termed a “seat-belt”, wraps around the CGa subunit to stabilize non-covalent association of the subunits, and also confers receptor selectivity (5). Structure and charge of the three N-linked carbohydrate chains influence activity; the most complex forms have lower activity but a longer halflife (1, 5). Bovine and porcine TSH bind human TSH receptors (TSHR) with high affinity (6). The hypothalamic peptide TRH stimulates production and secretion of TSH by thyrotrophs (basophile cells) in the anterior pituitary gland (1). TSH travels to thyroid TSHR to stimulate production of thyroxine (T4) (1). In the tissues, T4 is converted to the active form of thyroid hormone, triiodothyronine (T3), which completes a feedback loop by inhibiting TSH production (1, 7). Studies in the mouse identify bone marrow as a secondary site of TSH production (8). In bone, TSH signaling through TSHR on osteoblast and osteoclast precursors negatively regulates skeletal remodeling (9, 10). Bone marrow cells that produce TSH may also circulate to the thyroid and appear to modulate thyroid hormone activity in times of immunological stress (7, 11).

1. Szkudlinski, M.W. et al. (2002) Physiol. Rev. 82:473.
2. Sairam, M.R. and C.H. Li (1977) Can. J. Biochem. 55:755.
3. Fiddes, J.C. and H.M. Goodman (1979) Nature 281:351.
4. Wondisford, F.E. et al. (1988) J. Biol. Chem. 263:12538.
5. Weintraub, B.D. and M.W. Szkudlinski (1999) Thyroid 9:447.
6. Nunez, M.R. et al. (2004) Thyroid 14:991.
7. Matsushita, A. et al. (2007) Mol. Endocrinol. 21:865.
8. Klein, J.R. and H-C. Wang (2004) J. Exp. Biol 207:55.
9. Abe, E. et al. (2003) Cell 115:151.
10. Sun, L. et al. (2006) Ann. N.Y. Acad. Sci. 1068:309.
11. Klein, J.R. (2006) Exp. Biol. Med. 231:229.

Bioinformatics

• Uniprot:
  • P01215 (TSH alpha ) & P01222 (TSH beta )()