Recombinant Human ASGR1/ASGPR1 Fc Chimera Protein, CF

Catalog # Availability Size / Price Qty
10255-AS-050
Recombinant Human ASGR1/ASGPR1 Fc Chimera Protein, CF Bioactivity
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Recombinant Human ASGR1/ASGPR1 Fc Chimera Protein, CF Summary

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
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. When Recombinant Human ASGR1/ASGPR1 Fc Chimera (Catalog # 10255-AS) is immobilized at 2.5 µg/mL (100 µL/well) on a Goat Anti-Human IgG Fc antibody (Catalog # G-102-C) coated plate, the concentration of biotinylated beta -Gal-NAc-PAA that produces 50% of the optimal binding response is 0.75‑6 μg/mL. Measured by its binding ability in a functional ELISA. Immobilized Recombinant Human ASGR1/ASGPR1 Fc Chimera (Catalog # 10255-AS) binds human plasma von Willebrand Factor with an ED50 of 0.15-0.9 μg/mL.
Source
Human embryonic kidney cell, HEK293-derived human ASGR1/ASGPR1 protein
MDHuman IgG1
(Pro100-Lys330)
IEGRHuman ASGR1
(Asn63-Leu291)
Accession # P07306-1
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis

Met

Structure / Form
Disulfide-linked homodimer
Predicted Molecular Mass
53 kDa
SDS-PAGE
59-71 kDa, under reducing conditions

Product Datasheets

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.

10255-AS

Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 500 μg/mL in 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.

Data Images

Bioactivity Bioactivity View Larger

Immobilized Recombinant Human ASGR-1/ASGPR1 Fc Chimera (Catalog # 10255-AS) binds human plasma von Willebrand Factor with an ED50 of 150‑900 ng/mL.

SDS-PAGE SDS-PAGE View Larger

2 μg/lane of Recombinant Human ASGR1/ASGPR1 Fc Chimera (Catalog # 10255-AS) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 59-71 kDa and 120-140 kDa, respectively.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ASGR1/ASGPR1

The human asialoglycoprotein receptor (ASGPR) is an endocytic recycling receptor that belongs to the long-form subfamily of the C-type/Ca+2-dependent lectin family (1, 2, 3). It is a complex of two noncovalently-linked subunits, a major 46 kDa glycoprotein (ASGR1) and a minor 50 kDa glycoprotein (ASGR2). The major human ASGPR subunit, ASGR1 (also H1), is synthesized as a 291 amino acid (aa) type II transmembrane (TM) glycoprotein. It contains a 40 aa cytoplasmic region, a 21 aa TM segment, and a 230 aa extracellular domain (ECD) (4 - 6). The cytoplasmic region contains one palmitoylation site at Cys36 that is essential for ligand endocytosis and dissociation (7). The ECD contains two important structural regions. The first is a stalk region of 62 aa (aa 61-123) that contributes to noncovalent oligomerization. The second is a 118 aa, carbohydrate-binding, Ca+2-dependent C-type lectin domain (aa 161-278) that is stabilized by three Ca+2 ions (3, 8). Human ASGR1 ECD is 79% aa identical to mouse ASGR1 ECD. There are two minor (ASGR2) subunits that interact with ASGR1/H1 in a mutually exclusive manner to generate a functional ASGPR (9). They represent alternate splice forms of a type II TM protein. Termed H2b and H2c, H2b differs from H2c only by the presence of a 19 aa insert in its cytoplasmic region. This insert is significant because it allows serine phosphorylation of the cytoplasmic tail and provides for the majority of ASGPR ligand internalization (9). The stoichiometry of a functional ASGPR is unclear, but is suggested to be either a 2:2, 3:1 or 3:2 ratio of ASGR1/H1:ASGR2/H2 (9, 10, 11). ASGPR is found on hepatocytes and a subset of T cells (6, 12). ASGPR is reported to bind Gal (nonreducing), GalNAc, and sialic acid alpha 2,6Gal and GalNAc (3, 13, 14, 15). This is generally within the context of triantennary or tetraantennary configurations (2). The sialic acid terminations are of particular interest because molecules with these motifs most likely represent the endogenous ligands for ASGPR (14).

References
  1. Stockert, R. J. (1995) Physiol. Rev. 75:591.
  2. Weigel, P.H. and J.H.N. Yik (2002) Biochim. Biophys. Acta 1572:341.
  3. Meier, M. et al. (2000) J. Mol. Biol. 300:857.
  4. Spiess, M. et al. (1985) J. Biol. Chem. 260:1979.
  5. Spiess, M. and H.F. Lodish (1986) Cell 44:177.
  6. Bischoff, J. et al. (1988) J. Cell Biol. 106:1067.
  7. Yik, J.H.N. et al. (2002) J. Biol. Chem. 277:40844.
  8. Monroe, R.S. and B.E. Huber (1994) Gene 148:237.
  9. Yik, J.H.N. et al. (2002) J. Biol. Chem. 277:23076.
  10. Bider, M.D. et al. (1996) J. Biol. Chem. 271:31996.
  11. Lodish, H. (1991) Trends Biochem. Sci. 16:374.
  12. Park, J-H. et al. (2006) Biotechnol. Lett. 28:1061.
  13. Westerlind, U. et al. (2004) Glyconj. J. 21:227.
  14. Park, E.I. et al. (2005) Proc. Natl. Acad. Sci. USA 102:17125.
  15. Park, E.I. et al. (2003) J. Biol. Chem. 278:4597.
Long Name
Asialoglycoprotein Receptor 1
Entrez Gene IDs
432 (Human); 11889 (Mouse); 24210 (Rat)
Alternate Names
ASGPR 1; ASGP-R 1; ASGPR; ASGPR1; ASGR1; asialoglycoprotein receptor 1; CLEC4H1; CLEC4H1ASGPR1; C-type lectin domain family 4 member H1; Hepatic lectin H1; HL-1; MHL1; RHL1

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