Source	E. coli-derived human AG-2/AGR2 protein Arg21-Leu175, with a C-terminal 6-His tag
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.
N-terminal sequence Analysis	Met-Arg21
Predicted Molecular Mass	19 kDa
SDS-PAGE	17-20 kDa, under reducing conditions
Activity	Measured by the ability of the immobilized protein to support the adhesion of PC-3 human prostate cancer cells. The ED50 for this effect is 0.3-1.8 µg/mL.

Recombinant Human AG-2/AGR2 His-tag Protein, CF Scientific Data Examples

	Recombinant Human AG-2/AGR2 His-tag Protein Bioactivity Recombinant Human AG-2/AGR2 His-tag (Catalog # 10326-AG) supports the adhesion of PC-3 human prostate cancer cells. The ED₅₀ for this effect is 0.03-0.18 μg/mL.
	Recombinant Human AG-2/AGR2 His-tag Protein SDS-PAGE 2 μg/lane of Recombinant Human AG-2 His-tag (Catalog # 10326-AG) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 17-20 kDa.

Formulation, Preparation and Storage

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.

10326-AG

Formulation	Lyophilized from a 0.2 μm filtered solution in Tris, NaCl, and TCEP.
Reconstitution	Reconstitute at 500 μg/mL in water.
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.

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: AG-2/AGR2

AG-2 (Anterior gradient protein 2; also AGR2, GOB4 and HPC8) is an 18‑21 kDa member of the Protein disulfide-isomerase (PDI) family of enzymes. PDIs participate in the formation of transient disulfide linkages with molecules which aid in protein folding. Mature human AG-2 is 155 amino acids (aa) in length (aa 21‑175) and has a dimeric thioredoxin fold structure (1) which contains one cysteine (Cys81). It is expressed strongly in secretory cell types, such as prostate epithelium, small intestine goblet, trachea and lung, and is also expressed in estrogen receptor-positive breast cancer cell lines (2). Although AG-2 possesses a signal sequence, it appears to be rarely secreted, instead being found in the ER due the presence of a KTEL ER retention signal at its C-terminus (3). Over aa 21‑175, human AG-2 shares 94% aa identity with mouse AG-2. Over expressed in colorectal cancer stem cells, AG-2 is a marker regulated by the canonical Wnt/ beta -catenin pathway, AG-2 levels combined with elevated levels of beta -catenin are shown to indicate lower survival prognosis (4). Further, Wnt11-mediated signaling pathway has been implicated in secreted AG-2's role to promote metastasis of colorectal cancer (5). In prostate cancer, marker RAD9A affects the availability of AG-2. When added to RAD9A-depleted PC-3 cells, addition of AG-2 restores cell migration and adhesion properties (6). Additionally, AG-2 has been linked to pituitary adenomas as a biomarker (7).

Item	Value
References	Patel, P. et al. (2013) ‎J. Mol. Biol. 425:929. Thompson, D. A. and Weigel, R. J. (1998) Biochem. Biophys. Res. Commun. 251:111. Gupta, A. et al. (2012) J. Biol. Chem. 287:4773. Lamichane, B. et al. (2019) Biochem. Biophys. Res. Commun. 515:600. Tian, S. et al. (2018) Experimental Cell Research. 364:198. Broustas, C. G. et al. (2018) Carcinogenesis. 40:164. Tohti, M. et al. (2017) Clinical Neurology and Neurosurgery. 154:19.
Long Name	Anterior Gradient 2 Homolog
Entrez Gene IDs	10551 (Human); 23795 (Mouse); 298961 (Rat)
Alternate Names	AG-2, AG2, AG2member 17, AGR2, GOB-4, HPC8, PDIA17, anterior gradient 2 homolog (Xenopus laevis), anterior gradient homolog 2 (Xenopus laevis), hAG-2, secreted cement gland homolog

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Recombinant Human AG-2/AGR2 His-tag Protein, CF

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