肝素酶(HPSE)活性蛋白说明书

Active Heparanase (HPSE)

[ PROPERTIES ]

Source: Prokaryotic expression. Host: E. coli

Residues: Pro400~Ile543

Tags: N-terminal His-tag

Purity: >97%

Endotoxin Level: <1.0EU per 1μg (determined by the LAL method).
Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.05% sarcosyl

and 5% trehalose. Applications: Cell culture; Activity Assays.
(May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 9.9

Predicted Molecular Mass: 17.6kDa

Accurate Molecular Mass: 20kDa as determined by SDS-PAGE reducing conditions.

[ USAGE ]

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/mL. Do not vortex

[ STORAGE AND STABILITY ]

Storage: Avoid repeated freeze/thaw cycles.

Store at 2-8ºC for one month.

Aliquot and store at -80ºC for 12 months.

Stability Test: The thermal stability is described by the loss rate. The loss ratewas determined by accelerated thermal degradation test, that is, incubate theprotein at 37ºC for 48h, and no obvious degradation and precipitation wereobserved. The loss rate is less than 5% within the expiration date underappropriate storage condition.

[ SEQUENCE ]

[ ACTIVITY ] 

Heparanase(HPA), also known as HPSE, is an enzyme that acts both at thecell-surface and within the extracellular matrix to degrade polymeric heparansulfate molecules into shorter chain length oligosaccharides. Besides, EpithelialCell Transforming Sequence 2 (ECT2) has been identified as an interactor of HPA,thus a binding ELISA assay was conducted to detect the interaction ofrecombinant human HPA and recombinant human ECT2. Briefly, HPA were dilutedserially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100µL were thentransferred to ECT2-coated microtiter wells and incubated for 2h at 37℃. Wellswere washed with PBST and incubated for 1h with anti-HPA pAb, then aspiratedand washed 3 times. After incubation with HRP labelled secondary antibody, wellswere aspirated and washed 3 times. With the addition of substrate solution, wellswere incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wellsand read at 450nm immediately. The binding activity of HPA and ECT2 was shownin Figure 1, and this effect was in a dose dependent manner.

[ IDENTIFICATION ]

Figure 3. Western Blot

Sample: Recombinant HPA, Human;

Antibody: Rabbit Anti-Human HPA Ab

[ IMPORTANT NOTE ]

The kit is designed for research use only, we will not be responsible for any issueif the kit was used in clinical diagnostic or any other procedures.

本产品仅供科研使用。不能用于人和动物治疗等其它临床诊断使用