Active Plasminogen (Plg)

纤溶酶原(Plg)活性蛋白

[ PROPERTIES ]

Source: Prokaryotic expression. Host: E. coli

Residues: Gln274~Cys560

Tags: N-terminal His-tag

Purity: >95%

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: 6.7

Predicted Molecular Mass: 33.3kDa

Accurate Molecular Mass: 38kDa as determined by SDS-PAGE reducing conditions. Phenomenon explanation:

The possible reasons that the actual band size differs from the predicted are as follows:

1. Splice variants: Alternative splicing may create different sized proteins from the same gene. 2. Relative charge: The composition of amino acids may affects the charge of the protein. 3. Post-translational modification: Phosphorylation, glycosylation, methylation etc. 4. Post-translation cleavage: Many proteins are synthesized as pro-proteins, and then cleaved to

give the active form. 5. Polymerization of the target protein: Dimerization, multimerization etc.

[ 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 rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

[ SEQUENCE ]

[ ACTIVITY ]

Plasmin is an important enzyme present in blood that degrades many blood plasma proteins, including fibrin clots. It is released as a zymogen called plasminogen (PLG) from the liver into the systemic circulation. Plasmin is a serine protease that acts to dissolve fibrin blood clots. Apart from fibrinolysis, plasmin proteolyses proteins in various other systems: It activates collagenases, some mediators of the complement system and weakens the wall of the Graafian follicle(leading to ovulation). It cleaves fibrin, fibronectin, thrombospondin, laminin, and von Willebrand factor. Plasmin, like trypsin, belongs to the family of serine proteases. Besides, Actin Beta (ACTb) has been identified as an interactor of PLG,thus a binding ELISA assay was conducted to detect the interaction of recombinant human PLG and recombinant human ACTb. Briefly, PLG were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to ACTb-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PLG pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution , wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of PLG and ACTb was shown in Figure 1, and this effect was in a dose dependent manner.

Figure 1. The binding activity of PLG with ACTb.

[ IDENTIFICATION ]

Figure 2. SDS-PAGE

Sample: Active recombinant Plg, Human

Figure 3. Western Blot

Sample: Recombinant Plg, Human;

Antibody: Rabbit Anti-Human Plg Ab

[ IMPORTANT NOTE ]

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