ELISA Kit for Heparanase (HPSE)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 83-94 89
EDTA plasma(n=5) 88-105 94
heparin plasma(n=5) 84-94 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heparanase (HPSE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Heparanase (HPSE) 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) 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) 95-104% 93-102% 84-101% 92-99%
EDTA plasma(n=5) 88-99% 98-105% 80-93% 78-97%
heparin plasma(n=5) 79-99% 83-90% 78-98% 78-101%

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
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
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
RPA711Mu02 Recombinant Heparanase (HPSE) Positive Control; Immunogen; SDS-PAGE; WB.
RPA711Mu03 Recombinant Heparanase (HPSE) Positive Control; Immunogen; SDS-PAGE; WB.
RPA711Mu04 Recombinant Heparanase (HPSE) Positive Control; Immunogen; SDS-PAGE; WB.
RPA711Mu01 Recombinant Heparanase (HPSE) Positive Control; Immunogen; SDS-PAGE; WB.
PAA711Mu01 Polyclonal Antibody to Heparanase (HPSE) WB; IHC; ICC; IP.
PAA711Mu04 Polyclonal Antibody to Heparanase (HPSE) WB; IHC; ICC; IP.
PAA711Mu03 Polyclonal Antibody to Heparanase (HPSE) WB; IHC; ICC; IP.
PAA711Mu02 Polyclonal Antibody to Heparanase (HPSE) WB; IHC; ICC; IP.
LAA711Mu71 Biotin-Linked Polyclonal Antibody to Heparanase (HPSE) WB; IHC; ICC.
SEA711Mu ELISA Kit for Heparanase (HPSE) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA711Mu Multiplex Assay Kit for Heparanase (HPSE) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.