Recombinant Human Amnionless Protein, CF

Catalog # Availability Size / Price Qty
1860-AM-050
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human Amnionless Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its binding ability in a functional ELISA. Immobilized rhCubulin at 1 µg/mL can bind rhAmnionless with an apparent
KD < 100 nM.
Source
Mouse myeloma cell line, NS0-derived human Amnionless protein
Val20-Ala358, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Val20
Predicted Molecular Mass
37.2 kDa
SDS-PAGE
41-42 kDa, reducing conditions

Product Datasheets

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1860-AM

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

1860-AM

Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
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Background: Amnionless

Amnionless (AMN) is an approximately 48 kDa type I transmembrane protein that is required for surface expression of the extracellular membrane-associated protein Cubilin (1 - 5). The two form a complex termed CUBAM (4, 5). The 453 amino acid (aa) human AMN precursor contains a 19 aa signal sequence, a 338 aa extracellular domain (ECD), a 21 aa transmembrane domain, and a 75 aa cytoplasmic domain. The ECD contains two potential N-glycosylation sites and a cysteine-rich vWFC domain. Two cytoplasmic consensus FxNPxP/F sequences for clathrin-coated pit targeting mediate endocytosis of compounds bound by Cubulin (3, 5). The ECD of human AMN shares 67%, 66%, 76% and 78% aa identity with mouse, rat, canine and bovine AMN, respectively. AMN is present during gastrulation in the visceral endoderm and required for primitive streak formation during embryonic development in mouse (6). Transcription from an alternate start site in humans results in an isoform starting at M55 that may also function in development (7). AMN is highly expressed on polarized epithelia in the apical brush border membranes of small intestine and kidney proximal convoluted tubules (3 - 5, 7). Intestinal CUBAM is required for absorption of cobalamin (vitamin B12) when complexed with intrinsic factor (IF), and mutations of AMN or Cubilin cause Imersund-Grasbeck syndrome, also called megaloblastic anemia-1 (4, 5, 7). In the kidney, the CUBAM complex is thought to be important for reabsorption of proteins from filtrate, notably albumin and the molecules it carries (3, 8). Cubilin or CUBAM can be tightly associated with megalin, another endocytic receptor that contributes to stable expression of Cubilin, and to uptake of vitamin and lipoprotein complexes in the kidney and placenta (1, 9).

References
  1. Kozyraki, R. and F. Gofflot (2007) Curr. Pharm. Des. 13:3038.
  2. Coudroy, G. et al. (2005) J. Am. Soc. Nephrol. 16:2330.
  3. Strope, S. et al. (2004) Development 131:4787.
  4. He, Q. et al. (2005) Blood 106:1447.
  5. Fyfe, J.C. et al. (2004) Blood 103:1573.
  6. Kalantry, S. et al. (2001) Nat. Genet. 27:412.
  7. Tanner, S.M. et al. (2003) Nat. Genet. 33:426.
  8. Birn, H. (2006) Am. J. Physiol. Renal Physiol. 291:F22.
  9. Ahuja, R. et al. (2008) Biochem. J. 410:301.
Entrez Gene IDs
81693 (Human); 93835 (Mouse)
Alternate Names
AMN; amnionless homolog (mouse); Amnionless; PRO1028; protein amnionless; UNQ513/PRO1028; visceral endoderm-specific type 1 transmembrane protein

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