Human Histidine-Rich Glycoprotein, HRG ELISA Kit

Sample

Plasma, tissue homogenates and other biological fluids.

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Human HRG/HPRG ELISA Pair Set is for the quantitative determination of Human HRG/HPRG.
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 HRG/HPRG monoclonal antibody (in PBS, pH 7.4). Dilute to a working concentration of 2 μg/mL in PBS before coating.
Detection Antibody: 0.2 mg/mL of rabbit anti-Human HRG/HPRG polyclonal antibody conjugated to horseradish-peroxidase (HRP) (in PBS, 50 % HRP-Protector, pH 7.4, store at 4°C). Dilute to working concentration of 0.4 μg/mL in detection antibody dilution buffer before use.
Standard: – Each vial contains 35 ng of recombinant Human HRG/HPRG. 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 1000 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 HRG/HPRG was determined to be approximately 15.63 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

Histidine-rich glycoprotein, also known as HRG and HPRG, is a glycoprotein located in plasma and platelets, and contains an unusually large amount of histidine and proline. In human, five distinct domains are recognized in the mature HPRG molecule. There are two N-terminal cystatin-like modules (aa 19-254) and one His-Pro-rich region (aa 350-497) that is flanked by two Pro-rich segments (aa 276-321 and 498-525). The His-Pro-rich region contains 10 tandem repeats with an HHPHG motif, and the Nand C-termini are linked by a disulfide bond. The specific functions of HRG remain unclear, but it is known that the protein binds heme, dyes and divalent metal ions. It inhibits rosette formation and interacts with heparin, thrombospondin and plasminogen. Two of the protein's effects, the inhibition of fibrinolysis and the reduction of inhibition of coagulation, indicate a potential prothrombotic effect. HPRG is evolutionarily, functionally and structurally related to cleaved high molecular weight kininogen (HKa), an anti-angiogenic polypeptide that stimulates apoptosis of proliferating endothelial cells through binding to cell-surface tropomyosin. The antiangiogenic activity of the multidomain plasma protein HPRG is localized to its histidine-proline-rich (H/P) domain and has recently been shown to be mediated, at least partially, through binding to cell-surface tropomyosin in fibroblast growth factor-2-activated endothelial cells.

Standard Curve