Human Hemopexin, HPX ELISA Kit

Sample

Plasma, tissue homogenates and other biological fluids.

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Human Hemopexin / HPX ELISA Pair Set is for the quantitative determination of Human Hemopexin / HPX.
This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs.

Contents of Kit

Capture Antibody: 1 mg/mL of mouse anti-Human Hemopexin / HPX monoclonal antibody (in PBS, pH 7.4). Dilute to a working concentration of 2 μg/mL in PBS before coating.
Detection Antibody: 0.4 mg/mL of rabbit anti-Human Hemopexin / HPX polyclonal antibody conjugated to horseradish-peroxidase (HRP) (in PBS, 50 % glycerol, pH 7.4). Dilute to working concentration of 0.5 μg/mL in detection antibody dilution buffer before use.
Standard: Each vial contains 48 ng of recombinant Human Hemopexin / HPX. Reconstitute with 1 mL detection antibody dilution buffer. After reconstitution, store at -20°C to -80°C in a manual defrost freezer. A seven-point standard curve using 2-fold serial dilutions in sample dilution buffer, and a high standard of 1200 pg/mL is recommended.

Storage

Capture Antibody: Aliquot and store at -20°C to -80°C for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.
Detection Antibody: Store at 4°C and protect it from prolonged exposure to light for up to 6 months from date of receipt. Do not freeze!
Standard: Store lyophilized standard at -20°C to -80°C for up to 6 months from date of receipt. Aliquot and store the reconstituted standard at -80°C for up to 1 month. Avoid repeated freezethaw cycles.

Sensitivity

The minimum detectable dose of Human Hemopexin / HPX was determined to be approximately 18.8 pg/ml.
This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.

General Description

Hemopexin (HPX) is a serum glycoprotein responsible for the transport of haem to tissues such as liver, by receptor-mediated endocytosis, in an analogous manner to the iron transport protein transferrin, with recycling of intact haemopexin. Hemopexin binds heme with the highest affinity of any known protein. Its performs functions in scavenging the heme released or lost by the turnover of heme proteins such as hemoglobin, protects the body from the oxidative damage that free heme can cause. In addition, hemopexin releases its bound ligand for internalisation upon interacting with a specific receptor situated on the surface of liver cells, thus implicated in preservation of the body's iron. High hemopexin levels indicate that there has been significant degradation of heme containing compounds and hemopexin is made to scavenge any heme it can, and high hemopexin levels are therefore one of the diagnostic features of a hemolytic anemia. Hemopexin is an acute-phase plasma glycoprotein, produced mainly by the liver and released into plasma, where it binds heme with high affinity. Other sites of hemopexin synthesis are the nervous system, skeletal muscle, retina, and kidney. The only known receptor for the heme-hemopexin complex is the scavenger receptor, LDL receptor-related protein (LRP)1, which is expressed in most cell types, thus indicating multiple sites of heme-hemopexin complex recovery. Hemopexin is a heme-binding plasma glycoprotein which, after haptoglobin, forms the second line of defense against hemoglobin-mediated oxidative damage during intravascular hemolysis. A decrease in plasma hemopexin concentration reflects a recent release of heme compounds in the extracellular compartment. Heme-hemopexin complexes are delivered to hepatocytes by receptor-mediated endocytosis after which hemopexin is recycled to the circulation. Altered serum or plasma concentrations of hemopexin are found not only in hemolytic anemias but also in other conditions such as chronic neuromuscular diseases and acute intermittent porphyria. The diagnostic utility of hemopexin is less clear in other pathological conditions. Further studies are necessary to fully establish the clinical significance of hemopexin determination.

Standard Curve