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ACTIVITY TEST
Buffer Formulation 20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300. Traits Freeze-dried powder Purity > 95% Isoelectric Point 6.8 Applications Cell culture; Activity Assays.
Figure. The binding activity of Slit2 with GREM1.
Slit is a family of secreted extracellular matrix proteins which play an important signalling role in the neural development of most bilaterians. Humans, mice and other vertebrates possess three Slit homologs: Slit1, Slit2 and Slit3. The Slit2 protein has recently been discovered to be associated with the development of new blood vessels from pre-existing vessels, or angiogenesis. Slit2 has been implicated in promoting angiogenesis in mice (both in vitro and in vivo), in the human placenta, and in tumorigenesis. Besides, Gremlin 1 (GREM1) has been identified as an interactor of Slit2, thus a binding ELISA assay was conducted to detect the interaction of recombinant rat Slit2 and recombinant rat GREM1. Briefly, Slit2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to GREM1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-Slit2 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 Slit2 and GREM1 was shown in Figure 1, and this effect was in a dose dependent manner.USAGE
Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.
STORAGE
Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.
STABILITY
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.
Image
Figure. Western Blot
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