cGMP(Cyclic GMP) ELISA Kit
环磷酸鸟苷(cGMP)酶联免疫吸附测定试剂盒
| 规格： | / 96T / 48T / 96T*5 |
| 价格： | / ￥2400 / ￥1650 / ￥10200 |
货号：E-EL-0083
反应性: 通用
检测范围: 0.78~50 pmol/mL
灵敏度: 0.47 pmol/mL
用途
该试剂盒用于体外定量检测血清、血浆或其他相关生物液体中cGMP浓度。
检测原理
本试剂盒采用竞争ELISA法。用cGMP抗原包被于酶标板上，实验时样品或标准品中的cGMP与包被的cGMP竞争生物素标记的抗cGMP单抗上的结合位点，游离的成分被洗去。加入辣根过氧化物酶标记的亲和素，生物素与亲和素特异性结合而形成免疫复合物，游离的成分被洗去。加入显色底物(TMB)，TMB在辣根过氧化物酶的催化下呈现蓝色，加终止液后变成黄色。用酶标仪在450nm波长处测OD值，cGMP浓度与OD450值之间呈反比，通过绘制标准曲线计算出样品中cGMP的浓度。
| 反应类型 | 竞争法 |
| 规格 | 96T |
| 反应时间 | 2.0h |
| 反应性 | 通用 |
| 检测方法 | Colormetric |
| 检测范围 | 0.78-50 pmol/mL |
| 灵敏度 | 0.47 pmol/mL |
| 样本体积 | 50μL |
| 样本类型 | 血清、血浆或其他相关生物液体 |
特异性
可检测样本中的cGMP，且与其它相关蛋白无明显交叉反应。
重复性
板内，板间变异系数均<10%。
典型数据
由于实验操作条件的不同（如操作者、移液技术、洗板技术和稳定条件等），标准曲线的OD值会有所差异。以下数据和曲线仅供参考，实验者需根据自己的实验建立标准曲线。
| (pmol/mL) | O.D | Average |
|---|---|---|
| 50 | 0.359 0.371 |
0.365 |
| 25 | 0.473 0.491 |
0.482 |
| 12.5 | 0.685 0.675 |
0.68 |
| 6.25 | 0.96 0.982 |
0.971 |
| 3.13 | 1.322 1.304 |
1.313 |
| 1.56 | 1.633 1.627 |
1.63 |
| 0.78 | 1.857 1.865 |
1.861 |
| 0 | 2.152 2.17 |
2.161 |
精密度
板内精密度:低浓度样本,中浓度样本和高浓度样本分别在1块板子上检测20次。
板间精密度:低浓度样本,中浓度样本和高浓度样本分别在3块板子上检测20次。
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pmol/mL) |
98.40 | 299.90 | 999.00 | 92.40 | 326.30 | 1055.70 |
| Standard
deviation |
5.20 | 13.50 | 35.00 | 4.80 | 18.60 | 53.80 |
| C V (%) | 5.28 | 4.50 | 3.50 | 5.19 | 5.70 | 5.10 |
回收率
分别往5个不同样本中添加已知浓度的目标蛋白，做回收实验，得出回收率范围和平均回收率。
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum (n=5) | 91-104 | 96 |
| EDTA plasma (n=5) | 91-108 | 99 |
| Cell culture media (n=5) | 86-97 | 91 |
线性
分别往5个样本中添加已知浓度的目标蛋白，做回收实验，得出回收率范围及平均回收率。将5个样本分别稀释2倍，4倍，8倍，16倍做回收实验，得出回收率范围及平均回收率。
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 88-100 | 89-102 | |
| Average (%) | 96 | 94 | 94 | |
| 1:4 | Range (%) | 94-107 | 88-101 | 89-101 |
| Average (%) | 101 | 94 | 95 | |
| 1:8 | Range (%) | 96-111 | 90-106 | 89-102 |
| Average (%) | 102 | 97 | 97 | |
| 1:16 | Range (%) | 94-108 | 92-103 | 90-104 |
| Average (%) | 102 | 97 | 97 | |
试剂盒组成及保存
未拆封的试剂盒可在4℃保存一周；如果一周以后才使用试剂盒，请拆开试剂盒按照下表中的条件分别保存各组分。
| 中文名称 | 英文名称 | 规格 | 保存条件 |
|---|---|---|---|
| ELISA酶标板(可拆卸) | Micro ELISA Plate(Dismountable) | 8 孔×12 条 | -20℃，可存放6个月 |
| 冻干标准品 | Reference Standard | 2支 | |
| 浓缩生物素化抗体(100×) | Concentrated Biotinylated Detection Ab | 1支 120 μL | |
| 浓缩HRP酶结合物(100×) | Concentrated HRP Conjugate | 1支 120 μL | -20℃(避光)，可存放6个月 |
| 标准品&样品稀释液 | Reference Standard & Sample Diluent | 1瓶 20 mL | 4℃.可存放6个月 |
| 生物素化抗体稀释液 | Biotinylated Detection Ab Diluent | 1瓶 14 mL | |
| 酶结合物稀释液 | HRP Conjugate Diluent | 1瓶 14 mL | |
| 浓缩洗涤液(25×) | Concentrated Wash Buffer (25×) | 1瓶 30 mL | |
| 底物溶液(TMB) | Substrate Reagent | 1瓶 10 mL | 4℃(避光) |
| 反应终止液 | Stop Solution | 1瓶 10 mL | 4℃ |
| 封板覆膜 | Plate Sealer | 5 张 | |
| 产品说明书 | Product Description | 1 份 | |
| 质检报告 | Certificate of Analysis | 1 份 |
说明：所有试剂瓶盖须旋紧以防止蒸发和微生物的污染。
试剂体积以实际发货版说明书为准。相关试剂在分装时会比标签上标明的体积稍多一些，请在使用时量取而非直接倒出。
试验所需自备物品
- 1.酶标仪(450nm波长滤光片)
- 2.高精度移液器，EP管及一次性吸头：0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL
- 3.37℃恒温箱，双蒸水或去离子水
- 4.吸水纸
操作步骤
|
|
1. 对应板孔中加入50μL标准品工作液或样本后，立即每孔加入50μL生物素化抗体工作液，37℃孵育90分钟 |
| 2. 弃掉板内液体，洗板3次 | |
| 3. 每孔加入100μL HRP酶结合物工作液，37℃孵育30分钟，弃掉板内液体，洗板5次 | |
| 4. 每孔加入90μL底物溶液，37℃孵育15分钟 | |
| 5. 每孔加入50μL终止液 | |
| 6. 立即在450nm波长下读数，处理数据 |
客户发表文献
- 文献: Sildenafil, a Type-5 Phosphodiesterase Inhibitor, Fails to Reverse Myeloid-Derived Suppressor Cell-Mediated T Cell Suppression in Cells Isolated From Tuberculosis Patients
样品类型: MDSC - 文献: Nucleotide de novo synthesis increases breast cancer stemness and metastasis via cGMP-PKG-MAPK signaling pathway
样品类型: Cell sample - 文献: Effect of bisphenol F on sexual performance and quality of offspring in Male Wistar rats
样品类型: Penile - 文献: Glutamine restores testicular glutathione-dependent antioxidant defense and upregulates NO/cGMP signaling in sleep deprivation-induced reproductive dysfunction in rats
样品类型: Penile - 文献: Inhibition of phosphodiesterase-4 in the spinal dorsal horn ameliorates neuropathic pain via cAMP-cytokine-Cx43 signaling in mice
样品类型: spinal cord - 文献: Inhibition of Phosphodiesterase 5 Promotes the Aromatase-Mediated Estrogen Biosynthesis in Osteoblastic Cells by Activation of cGMP/PKG/SHP2 Pathway
样品类型: UMR106 cell - 文献: Inhibitory effect of (pro)renin receptor decoy inhibitor PRO20 on endoplasmic reticulum stress during cardiac remodeling
样品类型: cardiomyocytes (CMs),cardiac fibroblasts (CFs) - 文献: Ajuga bracteosa Exerts Antihypertensive Activity in l-NAME-Induced Hypertension Possibly through Modulation of Oxidative Stress, Proinflammatory Cytokines, and the Nitric Oxide/Cyclic Guanosine Monophosphate Pathway
样品类型: serum - 文献: Bee bread mitigates downregulation of steroidogenic genes, decreased spermatogenesis, and epididymal oxidative stress in male rats fed with high-fat diet
样品类型: serum,testis - 文献: Ziziphus Oxyphylla hydro-methanolic extract ameliorates hypertension in L-NAME induced hypertensive rats through NO/cGMP pathway and suppression of oxidative stress related inflammatory biomarkers.
样品类型: Serum - 文献: Protective effect of the combination of essential oil from patchouli and tangerine peel against gastric ulcer in rats
样品类型: serum - 文献: Orlistat reverses intratesticular lactate transport decline and infertility in male obese rats
样品类型: testis - 文献: Osthole prevents acetaminophen-induced liver injury in mice
样品类型: Tissue homogenate - 文献: Prenatal chronic stress impairs the learning and memory ability via inhibition of the NO/cGMP/PKG pathway in the Hippocampus of offspring
样品类型: hippocampal tissue - 文献: Osthole prevents tamoxifen-induced liver injury in mice
样品类型: Tissue homogenate - 文献: L-Arginine reverses maternal and pre-pubertal codeine exposure-induced sexual dysfunction via upregulation of androgen receptor gene and NO/cGMP signaling
样品类型: penile - 文献: Dexlansoprazole prevents pulmonary artery hypertension by inhibiting pulmonary artery smooth muscle cell to fibroblast transition.
样品类型: plasma - 文献: Aggravated endothelial endocrine dysfunction and intimal thickening of renal artery in high-fat diet-induced obese pigs following renal denervation
样品类型: Tissue homogenate - 文献: C-Type Natriuretic Peptide Ameliorates Lipopolysaccharide-Induced Cardiac Dysfunction in Rats with Pulmonary Arterial Hypertension
样品类型: plasma - 文献: Simultaneous inhibition of neprilysin and activation of ACE2 prevented diabetic cardiomyopathy
样品类型: heart tissue，plasma - 文献: Effect of C-Type Natriuretic Peptide (CNP) on Spermatozoa Maturation in Adult Rat Epididymis
样品类型: epididymal tissue - 文献: FXYD3 promotes the proliferation, migration, and invasion of pancreatic cancer cells by regulating the cGMP-PKG signaling pathway
样品类型: pancreatic tissue,BxPC-3 cell,CAPAN2 cell - 文献: Effect of low androgen status on the expression of adenosine A2A and A2B receptors in rat penile corpus cavernosum
样品类型: corpus cavernosum - 文献: Sildenafil augments fetal weight and placental adiponectin in gestational testosterone-induced glucose intolerant rats
样品类型: Placenta - 文献: Sildenafil ameliorates leptin resistance and normalizes lipid handling in the hypothalamic and adipose tissues of testosterone-exposed pregnant rats
样品类型: plasma
