Human Transferrin Antibody

Catalog # Availability Size / Price Qty
AF2914-100
AF2914-SP
Human Transferrin Antibody in Western Blot
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Human Transferrin Antibody Summary

Species Reactivity
Human
Specificity
Detects Human Transferrin in ELISAs. Detects human Transferrin and Apo-Transferrin in Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Human plasma-derived
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 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.

Data Example

Western Blot Detection of Human Transferrin and Human Apo-Transferrin by Western Blot. View Larger

Detection of Human Transferrin and Human Apo-Transferrin by Western Blot. Western blot shows purified holo-transferrin, purified apo-transferrin, and human plasma. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human Transferrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2914) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Transferrin and Apo-Transferrin at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: Transferrin

Human Transferrin (Tf) is a single chain, 80 kDa member of the anion-binding superfamily of proteins (1-5). It is a bilobed molecule that is the product of an ancient gene duplication event (1, 6). Transferrin is synthesized as a 698 amino acid (aa) precursor that is divided into a 19 aa signal sequence plus a 679 aa mature segment that contains 19 intrachain disulfide bonds. The crystal structure of Tf reveals a protein with two flanking 340 aa globular domains. Each are composed of a beta -sheet surrounded by series of alpha -helices (1, 7). The N- and C-terminal flanking regions (or domains) will bind ferric iron through the interaction of an obligate anion (usually bicarbonate) and four amino acids (His, Asp, and two Tyr) (7, 8). Apotransferrin (or iron-free) will initially bind one atom of iron at the C-terminus, and this is followed by subsequent iron binding by the N-terminus to form holotransferrin (diferric Tf) (8, 9). Through its C-terminal iron-binding domain, holotransferrin will interact with the type I Tf receptor (TfR) on the surface of cells where it is internalized into acidified endosomes. Iron dissociates from the Tf molecule within these endosomes, and is transported into the cytosol as ferrous iron. At physiological pH, iron-free Apotransferrin is not bound by TfR. But at acidic pH, such as exists in the endosome, Apotransferrin has considerable affinity for TfR. Thus, it remains bound to TfR and is recycled back to the cell surface where a neutral pH environment dissociates ligand from receptor. Each Tf molecule recycles 100-150 times during its lifetime (8-11). In addition to TfR, transferrin is reported to bind to cubulin, IGFBP3, microbial iron-binding proteins and liver-specific TfR2 (7, 12, 13, 14). Transferrin is variably glycosylated and the degree of sfialylation is suggestive of certain clinical conditions (15). Finally, Tf is highly allelic and the gene codominant, with many single aa changes noted. Three general forms are known, based on standard electrophoretic mobility. Fast Tf is known as transferrin B, slow transferrin is transferrin D, and the middle migrating transferrin is type/variant C, the most common (16, 17). Mature human TF is 73% aa identical to both mouse and rat Tf, and 68% and 71% aa identical to bovine and equine Tf, respectively.

References
  1. Brus, C.M. et al. (2001) Nat. Struct. Biol. 4:919.
  2. Schaeffer, E. et al. (1987) Gene 56:109.
  3. MacGillivray, R.T.A. et al. (1983) J. Biol. Chem. 258:3543.
  4. Yang, F. et al. (1984) Proc. Natl. Acad. Sci. USA 81:2752.
  5. Uzan, G. et al. (1984) Biochem. Biophys. Res. Commun. 119:273.
  6. Zak, O. et al. (2002) Biochemistry 41:7416.
  7. Gomme, P.T. and K. B. McCann (2005) Drug Discov. Today 10:267.
  8. Liu, R. et al. (2003) Biochemistry 42:12447.
  9. Pakdaman, R. et al. (1999) J. Mol. Biol. 293:1273.
  10. Hemadi, M. et al. (2004) Biochemistry 43:1736.
  11. Aisen, P. et al. (2001) Int. J. Biochem. Cell Biol. 33:940.
  12. Kozyraki, R. et al. (2001) Proc. Natl. Acad. Sci. USA 98:12941.
  13. Boulton, I.C. et al. (1998) Biochem. J. 334:269.
  14. Robb, A. and M. Wessling-Resnick (2004) Blood 104:4294.
  15. Landberg, E. et al. (1995) Biochem. Biophys. Res. Commun. 210:267.
  16. Gorg, A. et al. (1983) Hum. Genet. 64:222.
  17. Bean, P. and J.B. Peter (1994) Clin. Chem. 40:2078.
Entrez Gene IDs
7018 (Human)
Alternate Names
Beta-1 metal-binding globulin; DKFZp781D0156; EC 3.4.21; PRO1557; PRO2086; Serotransferrin; Siderophilin; TF; Transferrin

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