ELISA Kit for Histidine Rich Glycoprotein (HRG)

HPRG; HRGP; Histidine-Proline Rich Glycoprotein; Thrombophilia Due To Elevated HRG

Specificity

This assay has high sensitivity and excellent specificity for detection of Histidine Rich Glycoprotein (HRG).
No significant cross-reactivity or interference between Histidine Rich Glycoprotein (HRG) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Histidine Rich Glycoprotein (HRG) and the recovery rates were calculated by comparing the measured value to the expected amount of Histidine Rich Glycoprotein (HRG) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-96 85
EDTA plasma(n=5) 94-101 97
heparin plasma(n=5) 78-94 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Histidine Rich Glycoprotein (HRG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Histidine Rich Glycoprotein (HRG) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Histidine Rich Glycoprotein (HRG) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 84-99% 96-104% 94-101% 79-102%
EDTA plasma(n=5) 85-96% 95-103% 97-104% 93-101%
heparin plasma(n=5) 81-91% 97-105% 89-103% 80-96%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
J. Proteome Res. Identification and Confirmation of Differentially Expressed Fucosylated Glycoproteins in the Serum of Ovarian Cancer Patients Using a Lectin Array and LC–MS/MS ACS: pr300330z
Medical Oncology Evaluation of histidine-rich glycoprotein tissue RNA and serum protein as novel markers for breast cancer Springer: Source
Int J Cancer The nuclear bile acid receptor FXR controls the liver derived tumor suppressor histidine‐rich glycoprotein PubMed: 25363753
Nucleic Acid Therapeutics Pharmacokinetic and Pharmacodynamic Investigations of ION-353382, a Model Antisense Oligonucleotide: Using Alpha-2-Macroglobulin and Murinoglobulin Double-Knockout Mice Pubmed:27031383
United States Patent Application Compositions and Methods for Diagnosing Lung Cancer y2016:0077095.html
Hypertension Research Combination of serum histidine-rich glycoprotein and uterine artery Doppler to predict preeclampsia Pubmed:29416108
journal of histochemistry & cytochemistry Ultrastructural Localization of Histidine-rich Glycoprotein in Skeletal Muscle Fibers: Colocalization With AMP Deaminase Pubmed: 31880188
DOKLADY BIOCHEMISTRY AND BIOPHYSICS Personalized Approach to Determination of Histidine-Rich Glycoprotein and E-Cadherin in Supernatants of Immunocompetent Blood Cells and Breast Biopsy … Pubmed: 32342301
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