Vaspin Human ELISA

Regulatery Status: RUO
Type: Sandwich ELISA, Biotin-labelled antibody
Other Names Status: Visceral adipose-specific serpin, Serpin A12, Visceral adipose tissue-derived serine protease inhibitor, OL-64, SERPINA12
Species: Human
Catalog No Size
Product Catalog No: RD191097200R Pack Size: 96 wells (1 kit)

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Product Features

Visceral adipose tissue – derived serpin A12 (vaspin), also named OL-64, an adipocytokine, is structurally a member of the serine protease family. Serpins are the most diverse family of protease inhibitors. Their typical structural feature is the core domain composed from 3 beta-sheets and 9 alpha-helixes. The inhibitory activity of vaspin has not been described up to now, but its reactive site loop is typical for this proteinase family. Human Vaspin protein is composed of 395 amino acids and has a molecular weight of approximately 45.2 kDa and predicted pI 9.26. The cDNA was first isolated from white adipose tissue of Otsuka Long-Evans Tokushima Fatty (OLETF) rats. Vaspin mRNA expression is specific for visceral adipose tissues and it is also found circulating in the serum. The level of serum vaspin increased with age up to the peak of obesity, body weight and insulin resistance in OLETF rats and decreases with worsening of diabetes. Vaspin expression is missing in the diabetes-resistant lean rats, LETO, in comparison to OLEFT rats, animal model of metabolic syndrome. Expression was also absent in the subdermal, brown fatty tissue and other non-adipose tissues in OLEFT rats. These findings lead to the conclusion that the target tissue for insulin sensitising effect of vaspin is white adipose tissue. In humans, elevated serum concentration of vaspin is associated with obesity and impaired insulin sensitivity. In patients with type 2 diabetes the correlation between increased vaspin levels and BMI and decreased insulin sensitivity has not been observed. Vaspin expression decreased when diabetes worsened and its levels normalised when insulin or pioglitazone was administered. Gender differences in vaspin serum levels have been found in separate studies by two different authors. The low levels of vaspin in serum seem to be typical for lean subjects and athletes with long-term physical training. On the other hand, serum vaspin concentration increased in overweight people after they lost weight because of increased exercise. This paradox has been explained by the fact that serum vaspin level is differentially regulated in the non-active resting state and after exercise.

Technical Sheet / Info

Intended use

The RD191097200R Human Vaspin ELISA is a sandwich enzyme immunoassay for the quantitative measurement of human vaspin (visceral adipose tissue-derived serpin).

  • It is intended for research use only
  • The total assay time is less than 3.5 hours
  • The kit measures vaspin in serum and plasma (EDTA, heparin)
  • Assay format is 96 wells
  • Standard is recombinant protein based
  • Quality Controls are human serum based
  • Components of the kit are provided ready to use, concentrated or lyophilized

Clinical Application

  • Metabolic syndrome
  • Obesity
  • Insulin resistance

Test principle

In the Biovendor Human Vaspin ELISA, standards, quality controls and samples are incubated in microtitration wells pre-coated with polyclonal anti-human vaspin antibody. After a 60 minute incubation followed by washing, biotin labelled polyclonal anti-human vaspin antibody is added and incubated with the captured vaspin for 60 minutes. After another washing, streptavidin HRP conjugate is added. After 30 minutes incubation and the last washing step, the remaining conjugate is allowed to react with the substrate solution (TMB). The reaction is stopped by addition of acidic solution and absorbance of the resulting yellow product is measured. The absorbance is proportional to the concentration of vaspin. A standard curve is constructed by plotting absorbance values against vaspin concentrations of standards, and concentrations of unknown samples are determined using this standard curve.

Summary of protocol

  • Reconstitute QCs and Master Standard and prepare set of standards
  • Dilute samples 3×
  • Add 100 μl Standards, QCs and samples
  • Incubate at RT for 1 hour/300 rpm
  • Wash plate 5 times
  • Add 100 μl Biotin Labelled Antibody
  • Incubate at RT for 1 hour/300 rpm
  • Wash plate 5 times
  • Add 100 μl Streptavidin-HRP Conjugate
  • Incubate at RT for 30 min/300 rpm
  • Wash plate 5 times
  • Add 100 μl Substrate Solution
  • Incubate at RT for 10 min
  • Add 100 μl Stop Solution
  • Read absorbance and calculate results

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References

– Dimova R, Tankova T, Kirilov G, Chakarova N, Dakovska L, Grozeva G. Is vaspin related to cardio-metabolic status and autonomic function in early stages of glucose intolerance and in metabolic syndrome?. Diabetol Metab Syndr. 2016;8:46

– Kafalidis G, Boutsikou T, Briana DD, Boutsikou M, Marmarinos A, Baka S, Hassiakos D, Gourgiotis D, Malamitsi-Puchner A. Adipokines vaspin and omentin-1 are up-regulated in large for gestational age infants at term. Cytokine. 2013 Apr;62 (1):70-4

– Kiskac MM, Zorlu , Akkoyunlu ME, Kilic E, Karatoprak C, Cakirca M, Yavuz E, Ardic C, Camli AA, Cikrikcioglu M, Kart L. Vaspin and lipocalin-2 levels in severe obsructive sleep apnea. J Thorac Dis. 2014 Jun;6 (6):720-5

– Ostrowska Z, Ziora K, Oświęcimska J, Świętochowska E, Marek B, Kajdaniuk D, Strzelczyk J, Gołąbek K, Morawiecka-Pietrzak M, Wołkowska-Pokrywa K, Kos-Kudła B. Vaspin and selected indices of bone status in girls with anorexia nervosa. Endokrynol Pol. 2016;67(6):599-606. doi: 10.5603/EP.2016.0070

– Wu L, Piotrowski K, Rau T, Waldmann E, Broedl UC, Demmelmair H, Koletzko B, Stark RG, Nagel JM, Mantzoros CS, Parhofer KG. Walnut-enriched diet reduces fasting non-HDL-cholesterol and apolipoprotein B in healthy Caucasian subjects: a randomized controlled cross-over clinical trial. Metabolism. 2014 Mar;63 (3):382-91

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