Active Protein Kinase, cGMP Dependent Type II (PRKG2)

PRKGR2; cGKII; cGMP-dependent protein kinase 2

ACTIVITY TEST

Figure. The binding activity of PRKG2 with HSP90aA1.
Protein Kinase, cGMP Dependent Type II (PRKG2) belong to cGMP-dependent protein kinase or Protein Kinase G (PKG) which is a serine/threonine-specific protein kinase that is activated by cGMP. Two PKG genes, coding for PKG type I (PKG-I) and type II (PKG-II), have been identified in mammals. The PKG-I and PKG-II are homodimers of two identical subunits (~75kDa and ~85kDa, respectively) and share common structural features. PKG phosphorylates a number of biologically important targets and is implicated in the regulation of smooth muscle relaxation, platelet function, sperm metabolism, cell division, and nucleic acid synthesis. Besides, Heat Shock Protein 90kDa Alpha A1 (HSP90aA1) has been identified as an interactor of PRKG2, thus a binding ELISA assay was conducted to detect the interaction of recombinant human PRKG2 and recombinant human HSP90aA1. Briefly, PRKG2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100µL were then transferred to HSP90aA1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PRKG2 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 PRKG2 and HSP90aA1 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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Magazine Citations
PLoS One Enhanced heart failure, mortality and renin activation in female mice with experimental dilated cardiomyopathy pubmed:29240788
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