Vanin-1 Mouse/Rat ELISA Assay
The Vanin-1 Mouse/Rat ELISA Assay is For Research Use Only
Product Developed and Manufactured in Austria by Biomedica
Sensitivity: 2.31 pmol/l
Standard Range: 2.31 – 200 pmol/l
Incubation Time: 4.5 hours
Sample Type: Serum, plasma and urine
Sample Size: 5 µl / well
Conversion factor: 1 ng/ml = 19.2 pmol/l (MW: 52.07 kDa)
The Vanin-1 mouse/rat ELISA Assay is a sandwich enzyme immunoassay for the quantitative determination of mouse/rat Vanin-1 in serum, plasma or urine samples. In a first step, assay buffer is pipetted into the wells of the microtiter strips. Thereafter, standard/control/sample and detection antibody (polyclonal sheep anti-mouse Vanin-1-HRPO) are pipetted into the wells, which are pre-coated with anti-mouse Vanin-1 antibody. Vanin-1 present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody. In the washing step, all non-specific unbound material is removed. In a next step, the substrate (TMB, tetramethylbenzidine) is pipetted into the wells. The enzymecatalyzed color change of the substrate is directly proportional to the amount of Vanin-1 present in the sample. This color change is detectable with a standard microplate reader. A dose response curve of the absorbance (optical density, OD at 450 nm) versus standard concentration is generated using the values obtained from the standards. The concentration of Vanin-1 in the sample is determined directly from the dose response curve The kit utilizes recombinant mouse Vanin-1 as a calibrator. The VNN1 gene is conserved in chimpanzee, rhesus monkey, dog, cow, mouse, rat, and chicken.
Urine Vanin-1 ELISA Assay
Anti-Beta2 Glycoprotein 1 IgG ELISA
Vanin- (VAN1) is a GPI-anchored glycoprotein of 513 amino acids consisting of a base domain and anenzymatic nitrilase domain (Boersma et al., 2014). The ectoenzyme catalyzes the hydrolysis of pantetheine to pantothenic acid (vitamin B5) and cyteamine and thus, is involved in the regulation of oxidative stress and inflammation (Maras et al., 1999). Vanin-1 has a broad tissue expression with the highest levels being observed in kidney tubular epithelial cells (Pitari et al., 2000). The GPI anchor of Vanin-1 can be cleaved by a yet unknown mechanism, resulting in Vanin-1 being shed into the extracellular space.
Function: Vanin-1 is an epithelial ectoenzyme activating the conversion of pantetheine into pantothenic acid (vitamin B5) and cysteamine (Pitari et al., 2000). It has been suggested that the release of cysteamine by Vanin-1 promotes oxidative tissue damage and inflammation by inhibiting the activity of antioxidants like superoxide dismutase (SOD) and glutathione (GSH) (Hosohata et al., 2011; Saghaei et al., 2012). Indeed, Vanin-1 knockout mice have elevated stores of GSH and are more resistant to oxidative injury induced by whole-body gamma irradiation (Berruyer et al., 2004). On the other hand, several reports indicate that Vanin-1 might also act as tissue sensor for oxidative stress. In mice, antioxidant response-like elements could be identified in the promotor region of Vanin-1, which enhance the expression of Vanin-1 in the presence of oxidative stress (Berruyer et al., 2004). Similarly, Vanin-1 expression was shown to be upregulated in a human proximal tubular cell line after exposure to organic solvents (Hosohata et al., 2011). After renal ischemia-reperfusion in rats, a model involving oxidative tissue damage, renal Vanin-1 expression was also found to be upregulated (Yoshida et al., 2002). The highest levels of Vanin-1 expression could be assigned to renal tubular epithelial cells, while no expression is detectable in glomeruli (Hosohata et al., 2011; Pitari et al., 2000). Hence, Vanin-1 released from renal cells could be detectable in urine. In a study aimed to identify biomarkers for renal tubular injury, Hosohata and colleagues could indeed show in a rat model of nephrotoxicant-induced injury that Vanin-1 is upregulated in renal tubules earlier than other markers and shed into urine (Hosohata et al., 2011). Subsequent studies further verified the validity of Vanin-1 as an early biomarker of renal tubular damage in drug-induced acute kidney injury (Hosohata et al., 2012, 2016a), obstructive nephropathy (Washino et al., 2019) and hydronephrosis (Hosohata et al., 2018), diabetic nephropathy (Fugmann et al., 2011), renal injury in experimental colitis (Hosohata et al., 2014) and spontaneously hypertensive rats under high salt intake (Hosohata et al., 2016b; Washino et al., 2018). Of note, Vanin-1 seems to have superior predictive value for acute kidney injury than established markers KIM-1, NGAL, or NAG.