PKG (protein kinase G)

In a concentration-dependent manner, 8-Bromo-cGMP sodium (1-100 μM; 8 hours) can strengthen LLC-PK1 cells' resistance to CsA toxicity [3]. HO-1 protein synthesis is induced by 8-Bromo-cGMP sodium (1-100 μM; 16 hours) in a concentration-dependent manner [3].

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Using cultured proximal renal tubular epithelial cells (LLC-PK1), the present study investigates the effect of atrial natriuretic peptide (ANP) on cytotoxicity induced by cyclosporin A (CsA). Preincubation with ANP (1-100 nM) protected LLC-PK1 cells from CsA-induced toxicity in a concentration-dependent manner. A cytoprotective effect comparable to ANP was observed when preincubating the cells with 8-bromo cGMP (1-100 microM) or the antioxidant heme oxygenase (HO) metabolite bilirubin (0.1-10 microM). ANP or cGMP produced increases in HO-1 protein levels at concentrations that were also effective in cellular protection. Moreover, incubation with ANP or 8-Bromo-cGMP led to increased HO activity, i.e., formation of bilirubin in the cell lysate (up to 3-fold over basal). Tin protoporphyrin-IX (SnPP; 19 microM), an inhibitor of HO activity, completely abolished ANP-induced cytoprotection. Our results demonstrate that HO-1 is a cellular target of ANP and cGMP in renal cells. HO-1 induction and ensuing formation of antioxidant metabolites may be a novel pathway by which ANP protects from CsA-dependent nephrotoxicity and preserves renal function. [3]

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Protective effects of ANP and cGMP [3]
Incubation with CsA (48 h) resulted in a marked cytotoxicity and reduction of cell viability. Preincubation with ANP at 1–100 nM (8 h) diminished CsA toxicity in a concentration-dependent fashion and increased the surviving cell fraction by up to 70% (Fig. 1). Cytoprotection by ANP was less pronounced after shorter preincubation times and not detectable when CsA and ANP were added simultaneously to the cells (not shown). A similar cytoprotective effect was observed with the membrane permeable cGMP analog 8-Bromo-cGMP (1–100 μM). 8-Bromo cGMP at 1–100 μM increased resistance of LLC-PK1 cells to CsA toxicity concentration-dependently and augmented cell viability by up to 65% (Fig. 2). ANP and 8-bromo cGMP alone had no significant effect on cell viability under these conditions (not shown).

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ANP and cGMP increase HO activity and expression [3]
Cells were exposed to ANP or 8-Bromo-cGMP for 16 h. HO activity was assessed in the cell lysate by measuring formation of the HO metabolite bilirubin. ANP (0.1–1 μM) produced a concentration-dependent increase in HO activity up to 2.9-fold over basal levels (Fig. 5). Similar results were obtained when cells were incubated with 8-Bromo-cGMP (0.1–100 μM). A concentration-dependent stimulation of HO activity with a maximal 2.4-fold increase was detected in the presence of 8-bromo cGMP (Fig. 6). Stimulations of HO activity corresponded to increases in HO-1 protein synthesis Fig. 7, Fig. 8. ANP (1–100 nM) and 8-Bromo-cGMP (1–100 μM) induced the synthesis of HO-1 protein in a concentration-dependent fashion and produced maximal 3.1-fold and 3.7-fold elevations of basal HO-1 protein expression, respectively Fig. 7, Fig. 8.

In mice treated with vincristine up to the first hour of vehicle treatment, tail flick latency was significantly and dose-dependently increased by sodium 8-Bromo-cGMP (0.3, 1, 3.0 nmol; intrathecally; 10 min before testing). levels seen in male ICR mice weighing 20 g at 4 weeks of age. Vincristine can induce excruciating neuropathy in mice when administered at a dose of 0.05 mg/kg one day after tail flick latency and then 0.125 mg/kg twice a week for six weeks [4]. On a C57BL/6 background (19–35 g), 8-Bromo-cGMP sodium (10 mg/kg; i.v.; single dose) causes vasodilatory responses in WT littermates and eNOS-Tg mice [5].

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The rat formalin assay was used to assess effects of the cyclic guanosine mono-phosphate (cGMP) analog, 8-Bromo-cGMP on nociception and cGMP dependent protein kinase I (protein kinase G; PKG-I) expression in lumbar spinal cord. Intrathecal (i.t.) delivery of low doses of 8-bromo-cGMP (0.1–0.25 μmol) reduced nociceptive behavior and formalin-induced upregulation of PKG-I in the spinal cord. Medium doses (0.5–1 μmol i.t.) had no effect and high doses (2.5 μmol i.t.) caused hyperalgesia associated with a further increase of PKG-I expression and a PKG-I clip. To explain these dose-dependent contrary effects we assessed the potential involvement of various cGMP targets: protein kinase G, cyclic nucleotide gated cation channels (CNGs), phosphodiesterases (PDE2 and PDE3) and AMPA-receptors. The PKG inhibitor, Rp-8-bromo-cGMPS did not antagonize the antinociceptive effects of 8-Bromo-cGMP but caused antinociception itself. Inhibitors of CNGs, PDE2 and PDE3 had no effect on formalin evoked nociceptive behavior. S-AMPA however, antagonized the antinociceptive effects of 8-Bromo-cGMP. Since AMPA receptor currents were found to be reduced by 8-bromo-cGMP in vitro a direct or indirect reduction of AMPA receptor currents might possibly contribute to the antinociceptive effects of 8-bomo-cGMP. On the other hand, 8-bromo-cGMP evoked antinociception appears to be largely independent of PKG-I, CNGs, PDE2 and PDE3. The antinociceptive effects of the PKG inhibitor suggest that a strong PKG activation may be responsible for ‘high dose’ 8-Bromo-cGMP evoked hyperalgesia. [2]

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Nitric oxide (NO) plays a key role in regulating vascular tone. Mice overexpressing endothelial NO synthase [eNOS-transgenic (Tg)] have a 20% lower systemic vascular resistance (SVR) than wild-type (WT) mice. However, because eNOS enzyme activity is 10 times higher in tissue homogenates from eNOS-Tg mice, this in vivo effect is relatively small. We hypothesized that the effect of eNOS overexpression is attenuated by alterations in NO signaling and/or altered contribution of other vasoregulatory pathways. In isoflurane-anesthetized open-chest mice, eNOS inhibition produced a significantly greater increase in SVR in eNOS-Tg mice compared with WT mice, consistent with increased NO synthesis. Vasodilation to sodium nitroprusside (SNP) was reduced, whereas the vasodilator responses to phosphodiesterase-5 blockade and 8-Bromo-cGMP (8-Br-cGMP) were maintained in eNOS-Tg compared with WT mice, indicating blunted responsiveness of guanylyl cyclase to NO, which was supported by reduced guanylyl cyclase activity. There was no evidence of eNOS uncoupling, because scavenging of reactive oxygen species (ROS) produced even less vasodilation in eNOS-Tg mice, whereas after eNOS inhibition the vasodilator response to ROS scavenging was similar in WT and eNOS-Tg mice. Interestingly, inhibition of other modulators of vascular tone [including cyclooxygenase, cytochrome P-450 2C9, endothelin, adenosine, and Ca-activated K(+) channels] did not significantly affect SVR in either eNOS-Tg or WT mice, whereas the marked vasoconstrictor responses to ATP-sensitive K(+) and voltage-dependent K(+) channel blockade were similar in WT and eNOS-Tg mice. In conclusion, the vasodilator effects of eNOS overexpression are attenuated by a blunted NO responsiveness, likely at the level of guanylyl cyclase, without evidence of eNOS uncoupling or adaptations in other vasoregulatory pathways [5].

cGMP may either inhibit or facilitate synaptic transmission of nociceptive stimuli in the spinal cord. Hence, previously observed dual effects of NO are mirrored by dual effects of cGMP, where cGMP-induced hyperalgesia apparently involves PKG-I activation and upregulation whereas cGMP-induced antinociception is PKG independent. Since the antinociceptive effects require much less cGMP, antinociception appears to be the primary effect.[1]

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HO activity [3]
Confluent LLC-PK1 cells in 150-mm culture dishes were incubated for 16 h in the presence of control media, ANP, or 8-Bromo-cGMP. The method used for the determination of HO activity follows the protocol published by Motterlini and co-workers. Briefly, after the incubation, cells were washed twice with phosphate-buffered saline, gently scraped off the dish, and centrifuged (1000 × g for 10 min at 4°C). The cell pellet was suspended in MgCl2 (2 mM) phosphate (100 mM) buffer (pH 7.4), frozen at −70°C, thawed 3 times, and finally sonicated on ice before centrifugation at 18,000 × g for 10 min at 4°C. The supernatant (400 μl) was added to a NADPH-generating system containing 0.8 mM NADPH, 2 mM glucose-6-phosphate, 0.2 U glucose-6-phosphate-1-dehydrogenase, and 2 mg protein of rat liver cytosol prepared from the 105,000 × g supernatant fraction as a source of biliverdin reductase, potassium phosphate buffer (100 mM, pH 7.4), and hemin (10 μM) in a final volume of 200 μl. The reaction was conducted for 1 h at 37°C in the dark and terminated by addition of 1 ml chloroform. The extracted bilirubin was calculated by the difference in absorption between 464 and 530 nm using a quartz cuvette (extinction coefficient, 40 mM−1 × cm−1 for bilirubin). HO activity was measured as picomoles of bilirubin formed per milligram of endothelial cell protein per hour. Basal HO activity was in a range between 200 and 600 pmol bilirubin/mg protein/h.

Cell viability assay [3]
Cell Types: LLC-PK1 cells (ATCC CL 101)
Tested Concentrations: 1-100 μM
Incubation Duration: 8 hrs (hours)
Experimental Results: LLC-PK1 cells have increased tolerance to cyclosporine A (CsA) toxicity, And concentration-dependent and enhanced cell viability by up to 65%.

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Western Blot Analysis [3]
Cell Types: LLC-PK1 cells (ATCC CL 101)
Tested Concentrations: 1-100 μM
Incubation Duration: 16 hrs (hours)
Experimental Results: Induced the synthesis of HO-1 protein in a concentration-dependent manner.

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Cell viability analysis [3]
LLC-PK1 cells were seeded at 2 × 104 cells/well in 96-well microtiter plates in 100 μl of media containing 15% fetal bovine serum. After a 24-h incubation at 37°C, cells reached confluence and were incubated for 8 h in the presence of ANP, bilirubin, or 8-Bromo-cGMP. SnPP was added 10 min prior to ANP. Then, CsA was given to the cells without washing out the previously added agents. Incubation at 37°C was continued for 48 h, followed by a cytotoxicity assay. Cell viability was measured by staining with crystal violet as previously described. This colorimetric test allows assessment of the remaining viable cells after the incubation procedure. Cells were washed with phosphate-buffered saline, fixed with methanol for 5 min, and then stained for 10 min with a 0.1% crystal violet solution. Following 3 washes with tap water, the dye was eluted with 0.1 M trisodium citrate in 50% ethanol for 10 min. Optical density at 630 nm was measured using a microtiter plate reader.

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Western blot analysis [3]
LLC-PK1 were cultured in 150-mm dishes as described above. After a 16-h incubation with control media, ANP, or 8-Bromo-cGMP, cells were washed and extracted as described previously. Protein (100 μg) was applied to sodium dodecyl sulfate polyacrylamide gel electrophoresis. After electrophoresis, protein was transferred to a nitrocellulose membrane, and a polyclonal antibody to rat HO-1 was used to identify HO-1 protein content. Antigen antibody complexes were visualized with the horseradish peroxidase chemiluminescence system according to the manufacturer’s instructions. Quantitation of HO-1 protein content was performed using computer-assisted videodensitometry.

The rat formalin assay was used to assess effects of the cyclic guanosine mono-phosphate (cGMP) analog, 8-Bromo-cGMP on nociception and cGMP dependent protein kinase I (protein kinase G; PKG-I) expression in lumbar spinal cord. Intrathecal (i.t.) delivery of low doses of 8-bromo-cGMP (0.1–0.25 μmol) reduced nociceptive behavior and formalin-induced upregulation of PKG-I in the spinal cord. Medium doses (0.5–1 μmol i.t.) had no effect and high doses (2.5 μmol i.t.) caused hyperalgesia associated with a further increase of PKG-I expression and a PKG-I clip. To explain these dose-dependent contrary effects we assessed the potential involvement of various cGMP targets: protein kinase G, cyclic nucleotide gated cation channels (CNGs), phosphodiesterases (PDE2 and PDE3) and AMPA-receptors. The PKG inhibitor, Rp-8-bromo-cGMPS did not antagonize the antinociceptive effects of 8-bromo-cGMP but caused antinociception itself. Inhibitors of CNGs, PDE2 and PDE3 had no effect on formalin evoked nociceptive behavior. S-AMPA however, antagonized the antinociceptive effects of 8-Bromo-cGMP. Since AMPA receptor currents were found to be reduced by 8-bromo-cGMP in vitro a direct or indirect reduction of AMPA receptor currents might possibly contribute to the antinociceptive effects of 8-bomo-cGMP. On the other hand, 8-bromo-cGMP evoked antinociception appears to be largely independent of PKG-I, CNGs, PDE2 and PDE3. The antinociceptive effects of the PKG inhibitor suggest that a strong PKG activation may be responsible for ‘high dose’ 8-bromo-cGMP evoked hyperalgesia.[2]\n

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\n\nTo gain more insight into the role of cGMP in NO-induced hyperalgesia and antinociception we assessed effects of various doses of spinally delivered 8-Bromo-cGMP on nociceptive behavior and alterations of PKG-I expression in the spinal cord after formalin injection into the rat hind paw. In addition, we assessed the involvement of PKG-independent cGMP-targets including CNG channels, PDE2 and PDE3 and AMPA receptors. [2]
\nMale Sprague–Dawley rats (330–370 g) were used. Drugs were delivered through lumbar intrathecal (i.t.) catheters (12–13 cm polyethylene tubes) in a volume of 10 μl artificial cerebrospinal fluid (ACSF) followed by a 10 μl ACSF flush. Ten minutes later 50 μl of 5% formalin was injected subcutaneously into the dorsal surface of one hindpaw. Flinches were recorded for 60 min in 1 min intervals by a blinded observer. Rats were killed 1–96 h after formalin injection and the lumbar spinal cord was excised. [2]
\nWe first assessed the effects of the cGMP analog 8-Bromo-cGMP on the nociceptive behavior in the formalin assay and the expression of PKG-I in lumbar spinal cord (groups of 5–8 animals). We additionally assessed whether the effects of 8-bromo-cGMP (PKG-activator) were antagonized with Rp-8-bromo-cGMPS, a PKG inhibitor. In the second part of the study we investigated the potential involvement of cGMP regulated, but PKG independent pathways. Hence, we assessed the effects of the CNG channel inhibitor, L-cis-diltiazem (0.5 mg i.t.), the PDE2-inhibitor, erythro-hydroxy-nonyl-adenine (EHNA 0.25 μmol i.t.), the PDE3 inhibitor, milrinone (5 and 10 mg/kg i.p.) and effects of S-AMPA on 8-Bromo-cGMP-evoked effects (groups of four to six rats). The doses were chosen on the basis of previous studies and are at the upper tolerable range. Each part of the study included six control animals (ACSF i.t.).\n[2]

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\n\nAlterations in systemic vascular NO signaling. [5]
\nTo confirm previous observations that eNOS contributes to the lower vascular tone in the systemic bed of eNOS-Tg mice, we first studied the effect of the eNOS inhibitor l-NAME (100 mg/kg) in 10 WT and 10 eNOS-Tg mice. Subsequently, to study alterations in the NO signal transduction pathway, we determined the systemic vasodilator responses to the eNOS-dependent vasodilator acetylcholine (ACh, 200 μg/kg) (10 WT and 10 eNOS-Tg mice), the NO donor sodium nitroprusside (SNP, 300 μg/kg) (15 WT and 15 eNOS-Tg mice), the PDE5 inhibitor EMD-360527 (EMD, 30 mg/kg) (10 WT and 10 eNOS-Tg mice), or the PKG activator 8-Bromo-cGMP (8-Br-cGMP, 10 mg/kg) (10 WT and 10 eNOS-Tg mice).

[1]. Nitric oxide attenuates overexpression of Giα proteins in vascular smooth muscle cells from SHR: Role of ROS and ROS-mediated signaling. PLoS One. 2017 Jul 10;12(7):e0179301.

[2]. Dual effects of spinally delivered 8-bromo-cyclic guanosine mono-phosphate (8-bromo-cGMP) in formalin-induced nociception in rats. Neurosci Lett. 2002 Oct 31;332(2):146-50.

[3]. Atrial natriuretic peptide reduces cyclosporin toxicity in renal cells: role of cGMP and heme oxygenase-1. Free Radic Biol Med. 2002 Jan 1;32(1):56-63.

[4]. Possible involvement of the spinal nitric oxide/cGMP pathway in vincristine-induced painful neuropathy in mice. Pain. 2005 Sep;117(1-2):112-20.

[5]. Vasomotor control in mice overexpressing human endothelial nitric oxide synthase. Am J Physiol Heart Circ Physiol. 2007 Aug;293(2):H1144-53.

Sodium 8-bromo-3',5'-cyclic guanosine monophosphate is an organo-sodium salt with the counterion 8-bromoguanosine 3',5'-cyclic phosphate. It is a cell membrane-permeable cGMP analog that activates protein kinase G (PKG). Its PKG1α activation potency is 4.3 times that of cGMP, and it promotes relaxation of tracheal and vascular smooth muscle tissue in vitro. It acts as both a protein kinase G agonist and a muscle relaxant. It contains an 8-bromo-3',5'-cyclic guanosine monophosphate (1-) group.

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Decreased nitric oxide (NO) levels in vascular smooth muscle cells (VSMCs) of spontaneously hypertensive rats (SHR) may be the cause of Giα protein overexpression, which has been shown to be a contributing factor to the pathogenesis of hypertension in SHR. This study aimed to investigate whether increasing intracellular NO levels with the NO donor S-nitroso-N-acetyl-DL-penicillamine (SNAP) could attenuate the overexpression of Giα protein in vascular smooth muscle cells (VSMCs) of spontaneously hypertensive rats (SHR), and to explore its potential mechanism. Western blotting with specific antibodies was used to detect Giα protein expression and the phosphorylation levels of ERK1/2, growth factor receptors, and c-Src. After SNAP treatment of SHR VSMCs for 24 hours, the overexpression and proliferation of Giα-2 and Giα-3 proteins were significantly reduced, and this effect could not be reversed by the soluble guanylate cyclase inhibitor 1H(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one (ODQ). However, the MEK inhibitor PD98059 restored the SNAP-induced reduction in Giα protein expression to control levels. In addition, after SNAP treatment, the following indicators in SHR vascular smooth muscle cells (VSMCs) were restored to control levels: increased superoxide anion production, enhanced NAD(P)H oxidase activity, overexpression of AT1 receptor, Nox4, p22phox and p47phox proteins, increased levels of TBARS and protein carbonyl groups, and increased levels of PDGF-R, EGF-R, c-Src and ERK1/2 phosphorylation. These results indicate that NO reduces the overexpression and excessive proliferation of Giα-2/3 proteins in SHR VSMCs through a non-cGMP-dependent mechanism, and involves the transcriptional activation of ROS and its mediated EGF-R/PDGF-R and MAP kinase signaling pathways. [1]

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This study investigated the effect of atrial natriuretic peptide (ANP) on cyclosporine A (CsA)-induced cytotoxicity using cultured proximal renal tubular epithelial cells (LLC-PK1). Pre-incubation with ANP (1-100 nM) concentration-dependently protected LLC-PK1 cells from CsA-induced toxicity. Pre-incubation with 8-bromo-cGMP (1-100 μM) or bilirubin (0.1-10 μM), a metabolite of the antioxidant heme oxygenase (HO), showed comparable cytoprotective effects to ANP. ANP or cGMP increased HO-1 protein levels at concentrations that effectively protected cells. Furthermore, incubation with ANP or 8-bromo-cGMP increased HO activity, i.e., the production of bilirubin in cell lysates (up to 3 times the baseline level). The HO activity inhibitor tin psoralen IX (SnPP; 19 μM) completely eliminated the ANP-induced cytoprotective effect. Our results suggest that HO-1 is a target of ANP and cGMP in renal cells. HO-1 induction and the subsequent production of antioxidant metabolites may represent a novel pathway for ANP to protect the kidneys from CsA-dependent nephrotoxicity and maintain renal function. [3] The retrograde messenger NO is thought to either promote or inhibit synaptic transmission of nociceptive stimuli in the spinal cord. The choice between these two possibilities may depend on the amount of NO molecules released into the synaptic cleft. NO-induced hyperalgesia is thought to be mediated by activation of the cGMP/PKG pathway. As evidence, we found that high doses of the PKG activator cGMP analogue 8-bromo-cGMP induced hyperalgesia. However, low-dose intraspinal injection of 8-bromo-cGMP significantly reduced nociceptive behavior in the formalin test. This effect appears to be independent of PKG, as the PKG inhibitor Rp-8-Br-cGMPS not only failed to reverse the effect but also had an additive effect. Other potential cGMP targets, including CNG channels, PDE2, and PDE3, may also be unrelated to the analgesic effect induced by 8-bromo-cGMP, as relatively high doses of their respective antagonists—the CNG channel antagonist L-cis-diltiazem, the PDE2 antagonist EHNA, and the PDE3 antagonist milrinone—have no effect on noxious behavior. Recent studies have found that 8-bromo-cGMP can inhibit glutamate-induced AMPA receptor currents in hippocampal neurons, and this effect is independent of PKG activation, suggesting that 8-bromo-cGMP may directly or indirectly block AMPA receptors. In this study, intrathecal injection of S-AMPA completely eliminated the analgesic effect of “low-dose” 8-bromo-cGMP. Combined with previously reported in vitro results, it can be speculated that the reduction in AMPA receptor-induced excitatory postsynaptic potentials (EPSPs) may be one of the reasons why low-dose 8-bromo-cGMP exerts its analgesic effect. However, it remains unclear whether 8-bromo-cGMP directly targets AMPA receptors or nonspecifically reduces AMPA receptor-induced EPSP by inducing hyperpolarization. Therefore, the direct target of the analgesic effect of 8-bromo-cGMP remains unknown. As previous studies have shown, formalin treatment increases PKG-I protein levels in the spinal cord. This upregulation is observed at 1 hour and peaks at 48 hours. Both cGMP and cAMP have recently been identified as regulators of PKG gene expression in smooth muscle, but the regulatory mechanism of PKG expression in the spinal cord is unclear. High doses of 8-bromo-cGMP further enhanced PKG-I expression and resulted in two PKG bands in Western blot, one at the original PKG size of 73 kDa and the other at 65 kDa. Previous studies have shown that PKG can be cleaved into a catalytically active 65 kDa fragment that can be spontaneously activated without binding to cGMP. Therefore, Western blot results indicated that administration of high doses of 8-bromo-cGMP led to the cleavage of this PKG fragment. The resulting sustained and intense PKG activation may explain the significant increase in withdrawal behavior and nonspecific toxicity observed at high doses of 8-bromo-cGMP. In summary, our results suggest that cGMP may inhibit or promote synaptic transmission of nociceptive stimuli in the spinal cord. Thus, the previously observed dual effects of NO correspond to the dual effects of cGMP, where cGMP-induced hyperalgesia clearly involves the activation and upregulation of PKG-I, while cGMP-induced analgesia is independent of PKG. Since the amount of cGMP required for analgesia is much smaller, analgesia appears to be its primary function. [2]

C10H10BRN5NAO7P

446.08

444.939

C, 26.93; H, 2.26; Br, 17.91; N, 15.70; Na, 5.15; O, 25.11; P, 6.94

51116-01-9

31356-94-2 (Parent)

135419185

White to off-white solid powder

2.96 g/cm3

794.3ºC at 760 mmHg

434.2ºC

0.257

3

9

1

25

657

4

C1[C@@H]2[C@H]([C@H]([C@@H](O2)N3C4=C(C(=O)NC(=N4)N)N=C3Br)O)OP(=O)(O1)[O-].[Na+]

ZJRFCXHKYQVNFK-YEOHUATISA-M

InChI=1S/C10H11BrN5O7P.Na/c11-9-13-3-6(14-10(12)15-7(3)18)16(9)8-4(17)5-2(22-8)1-21-24(19,20)23-5;/h2,4-5,8,17H,1H2,(H,19,20)(H3,12,14,15,18);/q;+1/p-1/t2-,4-,5-,8-;/m1./s1

sodium;9-[(4aR,6R,7R,7aS)-7-hydroxy-2-oxido-2-oxo-4a,6,7,7a-tetrahydro-4H-furo[3,2-d][1,3,2]dioxaphosphinin-6-yl]-2-amino-8-bromo-1H-purin-6-one

8-Br-cGMP; 8BrcGMP; 8-Br-cGMP; 51116-01-9; 8-Br-cGMP; 8-Bromo-cGMP (sodium); 8-Bromoguanosine 3',5'-cyclic monophosphate sodium salt; 8-Bromo-cyclic GMP; CHEBI:64104; sodium 8-bromo-3',5'-cyclic GMP; MFCD00070128; 8-Bromo-cGMP (sodium); 8-Bromo-cyclic GMP; CHEBI:64104; sodium 8-bromo-3',5'-cyclic GMP; MFCD00070128; CID 16219005; 8 Br cGMP

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~100 mg/mL (~224.18 mM)

Solubility in Formulation 1: 100 mg/mL (224.18 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)