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​Product Spotlight: Kynurenine ELISA

August 16, 2017

IBL-America is featuring an ELISA for the quantitative determination of Kynurenine in serum or plasma. (split cursor)

The essential amino acid tryptophan is a constituent of proteins and is also a substrate for two important biosynthetic pathways: the generation of the neurotransmitter serotonin (5-hydroxytryptamine) and the formation of nicotinamide adenine dinucleotides via different kynurenine derivatives.
L-kynurenine is formed in the mammalian brain (40%) and is taken up from the periphery (60%), indicating that it can be transported across the blood-brain-barrier.
It can be converted by the inducible enzyme indoleamine 2,3-dioxygenase (IDO) into two other important compounds: the neuroprotective kynurenic acid and the neurotoxic quinolinic acid. The kynurenine pathway is also closely linked to the immune system, as different cytokines do influence the activity of the key enzymes.
An unbalanced tryptophan-kynurenine pathway is associated with different kind of diseases such as mental and psychotic disorders (e.g. depressions; Alzheimer´s disease; Huntington´s chorea), stress, and inflammatory neurological diseases (amyotrophic lateral sclerosis; ALS).

This Enzyme-linked Immunosorbent Assay (ELISA) is useful for measuring Kynurenine levels under different healthy and pathological conditions.

THIS NEW ELISA OFFERS THE FOLLOWING CHARACTERISTICS:

Product Name : Kynurenine ELISA
Catalog No. : IB89190

Description : Enzyme immunoassay (ELISA) for the quantitative determination of Kynurenine in serum and plasma samples

Sample Type/Volume : 20 µl serum or plasma

Design: Common sample preparation. Derivatization in liquid phase (48-well-reaction plate). Immunoassay using 12x8 break apart wells microtiter plate (precoated). 6 standards. 2 controls; ready for use.

Assay Time : Sample preparation and derivatization 30 min; ELISA overnight
Standard Range : 0/ 100 - 10 000 ng/ml
Sensitivity : 45,7 ng/ml
Storage : 2 - 8 °C
Regulatory Status : RUO
Kit size : 96 determinations

For more information: click here

Role of alpha Klotho in Chronic Kidney Disease, Metabolism, & Aging

August 9, 2017

Alpha Klotho is an approximately 130 kDa type I membrane protein that contains a short intracellular domain and two internal repeat regions (hKL1 and hKL2 in humans and mKL1 and mKL2 in mice) in its larger extracellular domain. We offer a sensitive alpha Klotho ELISA kit to study this protein target, which has been found to play a role in:

  • Chronic kidney disease (CKD)
  • Aging
  • Metabolism and calcium homeostasis
  • Cardiovascular disease
  • Inflammation

Expression of alpha Klotho

Alpha Klotho is preferentially expressed in parathyroid glands, the kidney distal convoluted tubules, and the brain choroid plexus (1). A soluble form of alpha Klotho is generated when the extracellular domain of membrane bound alpha-Klotho is shed and released into the circulation. Alternate splicing can also account for soluble alpha Klotho. The alpha Klotho protein has a global effect on organ health and function: mice lacking the klotho gene (kl/kl) show accelerated aging and numerous disease states, including osteoporosis, arteriosclerosis, skin atrophy, emphysema, pituitary gland abnormalities, Purkinje cell decrease and Parkinsonian gait, and atrophy of the genital organs and thymus (2).

Because alpha Klotho is involved in many organ systems, it is hypothesized that the protein is involved in fundamental metabolic processes. Consistent with this prediction, in homozygous kl /klmice blood glucose and insulin levels were significantly lower and insulin sensitivity was much higher compared to wild-type mice (3). In addition, soluble alpha Klotho helps regulate mineral metabolism, including circulating calcium and inorganic phosphate homeostasis (4). Manya et al. describe four alpha Klotho mediated pathways, including calcium channel activity, insulin/IGF signal inhibition, FGFR1(IIIc) binding and conversion to FGF23 receptor, and alpha1-Na+, K+-ATPase binding and recruitment of the Na+, K+-ATPase complex (1). Two of these Klotho pathways are illustrated below.

Measuring alpha Klotho

The Human soluble alpha-Klotho Assay (Cat.# 27998) uses a monoclonal antibody that has strong affinity and specificity for the tertiary protein structure of the alpha Klotho extracellular domain. The antibodies and substrates of the alpha Klotho ELISA were developed by Yamazaki and colleagues (4). This alpha Klotho ELISA can be used to accurately detect and measure human circulating alpha Klotho levels.

Our alpha Klotho Assay kit was compared with other commercially available kits for the measurement of alpha-Klotho in serum and plasma of patients with chronic kidney disease. In this experiment, the within- and between-run variations of the assay were <5% and <8%, respectively (5). These values were far better than the within-run variations of 32% and 13% noted for alpha Klotho assays purchased from Companies U and C, respectively.

This alpha Klotho assay kit has been used extensively to study the disease states and processes listed below:

Chronic Kidney Disease (CKD)

  • Alpha Klotho levels may be a useful indicator of kidney damage (6)
  • Lower serum alpha Klotho were associated with lower glomerular filtration rates and higher FGF23 levels (7)
  • Alpha Klotho was found to be significantly and transiently affected by cinacelcet treatment (8)

Aging

  • A link between senior citizen activities of daily living (ADLs) and alpha Klotho (9)
  • A heightened mortality risk may exist for individuals having lower circulating alpha Klotho levels (10)

Metabolism and Calcium Homeostasis

  • Fetal alpha-Klotho levels are significantly increased during certain gestational stages (11)
  • A possible link is found between alpha-Klotho and acromegaly (12)
  • Plasma alpha-Klotho levels are affected by abnormal nutritional states (13)

Cardiovascular Disease

  • Cardiovascular disease risk may be correlated with alpha-Klotho levels (14)
  • Low serum Klotho levels may correlate with poor vascular health (15)

Immunology and Inflammation

  • Alpha-Klotho may have inflammatory function (16)
References
1. Manya, H., et al. (2010) Geriatr. Gerontol. Int. 10 (Suppl. 1):S80.
2. Kuro-o, M., et al. (1997) Nature 390:45.
3. Utsugi, T., et al. (2000) Metabolism 49(9):1118-1123.
4. Yamazaki, Y., et al. (2010) Biochem. Biophys. Res. Commun. 398:513.
5. Heijboer, A.C., et al. (2013) Nephrol. Dial. Transplant. 0:1.
6. Pavik, I., et al. (2012) Clin. J. Am. Soc. Nephrol. 7:248.
7. Pavik, I., et al. (2013) Nephrol. Dial. Transplant. 28(2):352.
8. Komaba , H., et al.(2012) Nephrol. Dial. Transplant. 27(5):1967.
9. Semba, R.D., et al. (2011) J. Gerontol. A Biol. Sci. Med. Sci. 66A:794.
10. Crasto, C.L., et al. (2012) Rejuventation Res. 15(3):295.
11. Godang, K., et al. (2013) Eur. J. Endocrinol. 168:371.
12. Sze, L., et al. (2012) J. Int. Med. 272(1):93.
13. Amitani, M., et al. (2013) Nutrition 29(9):1106.
14. Semba, R.D., et al. (2011) J. Amer. Geriatrics Soc. 59(9):1596.
15. Kitagawa, M., et al. (2013). PLoS ONE 8(2): e56695.
16. Lam-Rachlin, J., et al. (2013) J. Perinatal Med. 0(0):1.

New Product Release | 3-Methoxytyramine RIA Kit

July 13, 2017

3-METHOXYTYRAMINE PLASMA RIA FAST TRACK (3-MT RIA) (IB88528)

ADVANTAGES:

  • New biomarker of phaeochromocytoma and paraganglioma tumours
  • The first commercial assay worldwide!
  • Speed and ease of use
  • 48 duplicate determinations or 96 single
  • Easy sample preparation (extraction)
  • No known interferences by medical drugs
  • High correlation with LC/MS-MS
  • Wide standard ranges, convenient measurement of pathological samples without predilution


COMPARISON LC-MS/MS: 3-Methoxytyramine RIA vs LC-MS/MS

ESSENTIAL INFORMATION

Cells derived from neuroendocrine tumors (e.g. pheochromcytoma) are known to produce catecholamines which are secreted episodically via vesicles into the bloodstream. Along with this, a small portion of the catecholamines is metabolized inside the cells to the corresponding catecholamines metabolites – namely metanephrine, normetanephrine and 3-methoxytyramine (3-MT) – which are secreted at low levels continuously into the bloodstream. 3-methoxytyramine is a metabolite of the neurotransmitter Dopamine formed by the introduction of a methyl group to dopamine by the enzyme catechol-O-methyl transferase (COMT). The measurement of 3-MT allows the detection of dopamine-producing tumors: For example a substantial number of subjects with head-and-neck paragangliomas (HNPGL) have biochemically active tumors and only display increased excretion of 3-MT, but not of other catecholamines or their metabolites. The combined levels of free metanephrine/normetanephrine and free 3-MT in plasma indicate a higher number of biochemically active HNPGLs than the 24-h urinary excretion rates of these markers. In addition, 3-MT acts as a neuromodulator and can also induce behavioral effects involved in movement control in a dopamine independent manner. Most of these effects are mediated by the trace amine associated receptor 1 (TAAR1) which in turn is involved in abnormal dopaminergic transmission, such as Parkinson's disease, dyskinesia and schizophrenia. The 3­-Methoxytyramine RIA is designed for research use only and should not be used in diagnostic procedures.


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New Product Release | Lipase Related Assays (HTGL / EL / GPIHBP1)

July 6, 2017

We have released lipase related assay kits (HTGL / EL / GPIHBP1) which can be measured with serum samples.

#27180 Human Serum HTGL ELISA kit
#27263 Human EL C-Terminal ELISA kit
#27179 Human GPIHBP1 ELISA kit

Please refer to the following articles as references.
HTGL
A new enzyme-linked immunosorbent assay system for human serum hepatic triglyceride lipase. Miyashita K et al. J Lipid Res. 2017 Jun 20. pii: jlr.M075432
PMID: 28634192
GPIHBP1
Autoantibodies against GPIHBP1 as a Cause of Hypertriglyceridemia. Beigneux AP et al. N Engl J Med. 2017 Apr 27;376(17):1647-1658.
PMID: 28402248


About HTGL
HTGL (hepatic triacylglycerol lipase / hepatic triglyceride lipase) is a secreted glycoprotein known as lipolytic enzyme or also hepatic lipase (HL). HTGL has an important role in lipoprotein metabolism as a lipolytic enzyme that hydrolyzes triglycerides (TG) and phospholipids in chylomicron remnants, intermediate-density lipoproteins (IDL) and high-density lipoproteins (HDL). It has been reported that hypercholesterolemia or triglyceridemia is observed and β-very-low-density lipoproteins (VLDL), chylomicron remnants, IDL, TG-rich low-density lipoproteins (LDL) and HDL are accumulated in patients with deficiency of HTGL.

About EL
EL (Endothelial Lipase) is a metabolic enzyme of HDL. It is a phospholipase A1 molecule that has a high substrate specificity against phospholipids that exists in high density lipoprotein, HDL molecules. It promotes HDL metabolism through metabolizing HDL phospholipids. It has been considered that EL is involved with lipid metabolism and elevating the risk of arteriosclerosis. Strong expression of EL on newly formed vessels (macrophages) in arteriosclerotic lesions is observed while EL expresses in vascular endothelial cells and vascular smooth myocytes of normal blood vessels. 

About GPIHBP1
GPIHBP1 (Glycosylphosphatidylinositol anchored high density lipoprotein binding protein 1) is an anchor protein that is modified by glycolipids and is known that it exists on a capillary endothelial cell membrane involved with metabolism of triglyceride rich (TG-rich) lipoprotein (triglyceride). GPIHBP1 transports lipoprotein lipase (LPL) that is synthesized and secreted by adipocyte or skeletal muscle cells into capillary lumen and moors on the surface of endothelial cells. It has been revealed that GPIHBP1 has a significantly important role in TG-rich lipoprotein metabolism as it has been considered that gene mutation of GPIHBP1 is a cause of type 1 hyperlipidemia (high chylomicronemia).

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New Drug Monitoring Assay | Vedolizumab (Entyvio®) (mAb-based) ELISA

June 26, 2017

We are pleased to introduce our latest addition to our Drug Monitoring Assay Line - Vedolizumab (Entyvio®) (mAb-based) ELISA!

The drug Vedolizumab (trade name Entyvio®) is a humanized immunoglobulin G1 monoclonal antibody that binds exclusively to the lymphocyte integrin α4β7. Identification of biomarkers might be beneficial for (non-) response and risk factors for adverse drug reactions that might be related to serum drug levels and maintaining the effective concentration in order to potentially avoid some side effects with a reliable method. The specificity of this testsystem is achieved by using a monoclonal antibody named "IG-19F3" for the coating of the microtiter plate. This antibody is specific for Vedolizumab only and does not cross react with other lymphocyte integrin α4β7 catchers.

Enzyme immunoassay for the specific determination of free Vedolizumab in serum and plasma. For research use only - not for use in diagnostic procedures.

Catalog NumberIG-AB116
SpeciesHuman
DesignEnzyme immunoassay (ELISA) technique.
Standards5 standards, ready to use.
ControlsNone provided.
Sample TypesSerum and plasma.
Sample Volume10 µL
Assay Desc.1 hour incubation (RT) + 30 min. (RT) + 20 min. (RT) = 1 hour, 50 min. total incubation time.
Standard Range0 / 6 - 200 ng/mL
Sensitivity5 ng/mL

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