Multiplex Assay Kit for Heparanase (HPSE) ,etc. by CBA (Cytometric Bead Array)

HPA; HPR1; HPSE1; HSE1; Endo-glucoronidase;

Specificity

This assay has high sensitivity and excellent specificity for detection of Heparanase (HPSE) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Heparanase (HPSE) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Heparanase (HPSE) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Heparanase (HPSE) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 89-104 95
EDTA plasma(n=5) 85-104 98
heparin plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heparanase (HPSE) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Heparanase (HPSE) ,etc. by CBA (Cytometric Bead Array) 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 Heparanase (HPSE) ,etc. by CBA (Cytometric Bead Array) 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) 81-99% 88-102% 84-105% 84-93%
EDTA plasma(n=5) 82-101% 86-93% 91-98% 82-105%
heparin plasma(n=5) 87-95% 86-101% 95-102% 94-105%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
Plos one PG545, a Heparan Sulfate Mimetic, Reduces Heparanase Expression In Vivo, Blocks Spontaneous Metastases and Enhances Overall Survival in the 4T1 Breast Carcinoma Model Plosone: Source
Journal of Medical Science Assessment of heparanase and heparin-binding growth and angiogenesis factors in the uterine cavity ? uid in women with impaired reproduction Nowinylekarskie:Source
Int J Clin Exp Pathol Unfractionated heparin attenuates intestinal injury in mouse model of sepsis by inhibiting heparanase PubMed: 26191183
Tumour Biology PI-88 inhibits postoperative recurrence of hepatocellular carcinoma via disrupting the surge of heparanase after liver resection PubMed: 26415733
Journal of Medical Science Assessment of heparanase and heparin-binding growth and angiogenesis factors in the uterine cavity fluid in women with impaired reproduction article:84
Renal Failure Plasma heparanase is associated with blood glucose levels but not urinary microalbumin excretion in type 2 diabetic nephropathy at the early stage pubmed:28994624
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