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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 > 90% Isoelectric Point 7.2 Applications Cell culture; Activity Assays.
Figure. The binding activity of SEMA3A with NRP1.
Semaphorin 3A (SEMA3A) is a member of the semaphorin family. Semaphorin 3A is secreted by neurons and surrounding tissue to guide migrating cells and axons in the developing nervous system. This secreted Sema3A protein can function as either a chemorepulsive agent, inhibiting axonal outgrowth, or as a chemoattractive agent, stimulating the growth of apical dendrites. In both cases, the protein is vital for normal neuronal pattern development. Besides, Neuropilin 1 (NRP1) has been identified as an interactor of SEMA3A, thus a binding ELISA assay was conducted to detect the interaction of recombinant human SEMA3A and recombinant human NRP1. Briefly, SEMA3A were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to NRP1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-SEMA3A 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 SEMA3A and NRP1 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.
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Figure. Gene Sequencing (Extract)
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Figure. SDS-PAGE
Figure. Western Blot
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