Haptoglobin ELISA Kit (HP)

Full Name: Haptoglobin ELISA Kit (HP)
Reactivity: Human
Sample Type: Plasma, Cell Supernatant, Serum
Sensitivity:0.156 ng/ml

INTRODUCTION

Haptoglobin (HP) is a polymorphic gene in humans, it is made of 2-alpha and 2-beta subunits. There subunits are found to vary amongst many different species and there is evidence to suggest different subtypes exist. It has be described as an immunoglobulin like plasma protein that is produced within the liver. One of its main functions of haptoglobin is the binding of free hemoglobin that have been release by RBCs. It is useful during the detection and evaluation of hemolytic anemia. Furthermore, plasma depletion is considered a useful biological marker in identifying the accelerated destruction of RBCs.

Haptoglobin gene mutations have been studied in depth for their influence on the levels in humans. Findings show that the A-allele is associated with low levels of serum haptoglobin, while a non-functional haptoglobin gene produces greater levels than normal. This functional difference can be found among different sexes and due to the presence or absence of certain mutations. Specifically, a functional haptoglobulin gene is mostly linked to increased circulating levels of hemolysed erythrocytes as well as anemia and sensitivity to oxidative stress.

There are many assays that have been used to analyse haptoglobin in both humans and animals and the most common is measuring the natural affinity via spectrophotometry. There are also immunoassay kits being used however these are not currently widely implemented. in the clinical setting. The natural affinity for hemoglobin is measured by a spectrophotometer and the extent of binding can be read at varying concentrations. The level in whole blood is determined by measuring the amount of haptoglobin in a centrifuged serum sample. There are currently no commercially available kits that measure levels of HpB1.

INTENDED USE

Human haptoglobin ELISA kit is ideal for analysing concentration of haptoglobin (HP) in human cell supernatant, plasma or serum samples.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• Microtitre Strips (96 Wells): Coated with mouse monoclonal antibody against haptoglobin.
• 10x Wash Buffer.
• 5x Assay Buffer.
• 100x Detection Antibody Buffer: Labelled biotin polyclonal antibody.
• Standard: Recombinant lyophilized.
• 200x STP-HRP Solution.
• Substrate Solution.
• Stop Solution.

TYPICAL RESULTS

For the following haptoglobin ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 3.12, 6.25, 12.5, 25, 50, 100 ng/ml.

ASSAY CHARACTERISTICS

– Sensitivity: 0.156 ng/ml
– Assay Range: 3.12 – 100 ng/ml.
– Specificity: No interference or cross reaction observed.
– Inter-Assay Precision: 1% (CV).
– Intra-Assay Precision: 5.63 (CV).
– Linearity: 95.37% (1:2 dilution), 98.1% (1:4 dilution), 110.82% (1:8 dilution).
– Other Names: Haptoglobin, Hp.

REFERENCES

1. Haptoglobin Therapeutics and Compartmentalization of Cell-Free Hemoglobin Toxicity. Trends Mol Med. (2020) 26 (7): 683-697. Buehler PW, et al.
2. The haptoglobin-CD163-heme oxygenase-1 pathway for hemoglobin scavenging. Oxid Med Cell Longev. (2013) 2013: 523652. Thomsen J.H., et al.
3. Hp: an inflammatory indicator in cardiovascular disease. Future Cardiol. (2016) 12 (4): 471-81. Graves K.L. and Vigerust D.J.
4. Biological functions of haptoglobin–new pieces to an old puzzle. Eur J Clin Chem Clin Biochem. (1997) 35 (9): 647-54. Dobryszycka W.

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