cAMP(Cyclic adenosine monophosphate) ELISA Kit
环磷酸腺苷(cAMP)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-0056
反应性: 通用
检测范围: 1.56~100 ng/mL
灵敏度: 0.94 ng/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cAMP浓度。
检测原理
本试剂盒采用竞争ELISA法。用cAMP抗原包被于酶标板上，实验时样品或标准品中的cAMP与包被的cAMP竞争生物素标记的抗cAMP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，cAMP浓度与OD450值之间呈反比，通过绘制标准曲线计算出样品中cAMP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 | 1.56-100 ng/mL |
| 灵敏度 | 0.94 ng/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cAMP，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。
| (ng/mL) | O.D | Average |
|---|---|---|
| 100 | 0.359 0.409 |
0.384 |
| 50 | 0.474 0.512 |
0.493 |
| 25 | 0.708 0.682 |
0.695 |
| 12.5 | 1.004 1.036 |
1.02 |
| 6.25 | 1.47 1.44 |
1.455 |
| 3.13 | 1.912 1.896 |
1.904 |
| 1.56 | 2.263 2.271 |
2.267 |
| 0 | 2.757 2.769 |
2.763 |
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 (ng/mL) |
5.40 | 14.10 | 37.40 | 5.90 | 15.00 | 34.90 |
| Standard
deviation |
0.30 | 0.70 | 2.00 | 0.40 | 0.90 | 1.80 |
| C V (%) | 5.56 | 4.96 | 5.35 | 6.78 | 6.00 | 5.16 |
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) | 89-104 | 95 |
| EDTA plasma (n=5) | 86-100 | 92 |
| Cell culture media (n=5) | 87-100 | 92 |
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 (%) | 85-100 | 86-98 | |
| Average (%) | 94 | 92 | 92 | |
| 1:4 | Range (%) | 91-103 | 87-101 | 101-114 |
| Average (%) | 97 | 92 | 107 | |
| 1:8 | Range (%) | 84-98 | 90-102 | 99-116 |
| Average (%) | 91 | 97 | 107 | |
| 1:16 | Range (%) | 86-100 | 90-102 | 99-114 |
| Average (%) | 92 | 97 | 105 | |
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: The gut microbe Bacteroides fragilis ameliorates renal fibrosis in mice
样品类型: renal,PRTC cells - 文献: Combining adoptive NK cell infusion with a dopamine-releasing peptide reduces senescent cells in aged mice
样品类型: Splenic NK cell - 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: Site-specific N-terminal PEGylation-based controlled release of biotherapeutics: An application for GLP-1 delivery to improve pharmacokinetics and prolong hypoglycemic effects
样品类型: HEK293 cell - 文献: Phosphodiesterase 4B activation exacerbates pulmonary hypertension induced by intermittent hypoxia by regulating mitochondrial injury and cAMP/PKA/p-CREB/PGC-1α signaling
样品类型: serum,pulmonary arterial smooth muscle cells (PASMCs) - 文献: Forskolin, an Adenylcyclase/cAMP/CREB Signaling Activator Restoring Myelin-Associated Oligodendrocyte Destruction in Experimental Ethidium Bromide Model of Multiple Sclerosis
样品类型: brain tissue - 文献: Muscone and (+)-Borneol Cooperatively Strengthen CREB Induction of Claudin 5 in IL-1β-Induced Endothelium Injury
样品类型: bEnd.3 cell - 文献: Ginsenoside Rg1 promotes neurite growth of retinal ganglion cells through cAMP/PKA/CREB pathways
样品类型: RGC-5s cell - 文献: Conjunctival Fluid Secretion Impairment via CaCC-CFTR Dysfunction Is the Key Mechanism in Environmental Dry Eye
样品类型: conjunctiva epithelium - 文献: A Novel Furocoumarin Derivative, 5-((diethylamino)me-13 thyl)-3-phenyl-7H-furo [3,2-g] chromen-7-one Upregulates Melanin Synthesis via the Activation of cAMP/PKA and MAPKs Signal Pathway: In Vitro and In Vivo Study
样品类型: skin - 文献: ONO-1301 loaded nanocomposite scaffolds modulate cAMP mediated signaling and induce new bone formation in critical sized bone defect
样品类型: Mesenchymal stem cell - 文献: Aspirin suppresses hepatic glucagon signaling through decreasing production of thromboxane A2
样品类型: Hepatocyte - 文献: Neuronal lack of PDE7a disrupted working memory, spatial learning, and memory but facilitated cued fear memory in mice
样品类型: hippocampus samples - 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: Anti-platelet aggregation of Panax notoginseng triol saponins by regulating GP1BA for ischemic stroke therapy
样品类型: Serum - 文献: CD38 affects the biological behavior and energy metabolism of nasopharyngeal carcinoma cells
样品类型: Cell lysate - 文献: Receptor-Mediated AKT/PI3K Signalling and Behavioural Alterations in Zebrafish Larvae Reveal Association between Schizophrenia and Opioid Use Disorder
样品类型: larvae - 文献: Pyruvate Upregulates Hepatic FGF21 Expression by Activating PDE and Inhibiting cAMP–Epac–CREB Signaling Pathway
样品类型: HepG2 cell - 文献: Developmental Exposure to Atrazine Impairs Spatial Memory and Downregulates the Hippocampal D1 Dopamine Receptor and cAMP-Dependent Signaling Pathway in Rats
样品类型: Tissue homogenate - 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: A2A R mediated modulation in IP3 levels altering the [Ca2+]i through cAMP-dependent PKA signalling pathway
样品类型: HEK 293 cell - 文献: The Inhibition by Guanfu Base A of Neuropathic Pain Mediated by P2Y12 Receptor in Dorsal Root Ganglia
样品类型: Cell culture supernatant - 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: The protective effect of the PDE-4 inhibitor rolipram on intracerebral haemorrhage is associated with the cAMP/AMPK/SIRT1 pathway
样品类型: Brain tissue - 文献: Structure-based redesigning of pentoxifylline analogs against selective phosphodiesterases to modulate sperm functional competence for assisted reproductive technologies
样品类型: sperm - 文献: cAMP agonist forskolin disrupts mitochondrial metabolism and induces senescence in human mesenchymal cells
样品类型: MSCs - 文献: Long Non-coding RNA Uc.48+ Small Interfering RNA Alleviates Neuroinflammatory Hyperalgesia in Gp120-Treated Rats via the P2Y12 Receptor
样品类型: HEK 293 cells - 文献: Effect of Natural Adenylcyclase/cAMP/CREB Signalling Activator Forskolin against Intra-Striatal 6-OHDA-Lesioned Parkinson’s Rats: Preventing Mitochondrial, Motor and Histopathological Defects
样品类型: brain - 文献: Serum Metabolomics Based on GC-MS Reveals the Antipyretic Mechanism of Ellagic Acid in a Rat Model
样品类型: cerebrospinal fluid (CSF) - 文献: Calcimimetic R568 improved cardiac remodeling by classic and novel renin-angiotensin system in spontaneously hypertensive rats:
样品类型: plasm - 文献: Morphine dependence and withdrawal-induced changes in mouse Sertoli cell (TM4) line: Evaluation of apoptotic, inflammatory and oxidative stress biomarkers
样品类型: TM4 cells - 文献: Osthole pretreatment alleviates TNBS-induced colitis in mice via both cAMP/PKA-dependent and independent pathways
样品类型: serum - 文献: Teasaponin suppresses Candida albicans filamentation by reducing the level of intracellular cAMP
样品类型: YEM30 cell - 文献: Analysis and application of a neutralizing linear epitope on liable toxin B of enterotoxin Escherichia coli
样品类型: epithelial cell - 文献: Osthole prevents tamoxifen-induced liver injury in mice
样品类型: Tissue homogenate - 文献: Curcumin restrains hepatic glucose production by blocking cAMP/PKA signaling and reducing acetyl CoA accumulation in high-fat diet (HFD)-fed mice
样品类型: liver,primary hepatocytes - 文献: Dopamine receptor D1 signaling stimulates lipolysis and browning of white adipocytes
样品类型: adipose tissue,adipocyte - 文献: Effect of phosphodiesterase-4 inhibitor rolipram on colonic hypermotility in water avoidance stress rat model
样品类型: colon tissue - 文献: Royal Jelly Alleviates Cognitive Deficits and β-Amyloid Accumulation in APP/PS1 Mouse Model Via Activation of the cAMP/PKA/CREB/BDNF Pathway and Inhibition of Neuronal Apoptosis
样品类型: Tissue homogenate - 文献: Eleutheroside E Enhances the Long-Term Memory of Radiation-Damaged C. elegans through G-Protein-Coupled Receptor and Neuropeptide Signaling Pathways
样品类型: Tissue homogenate - 文献: Biatriosporin D displays anti-virulence activity through decreasing the intracellular cAMP levels
- 文献: Down-regulation of A3AR signaling by IL-6-induced GRK2 activation contributes to Th17 cell differentiation
样品类型: Cell sample - 文献: Calcimimetic R568 reduced the blood pressure and improved aortic remodeling in spontaneously hypertensive rats by inhibiting local renin‑angiotensin system activity
样品类型: Plasma,Tissue homogenate - 文献: Calcilytic NPS2143 promotes proliferation and inhibits apoptosis of spontaneously hypertensive rat vascular smooth muscle cells via activation of the renin‑angiotensin system
样品类型: Cell lysate - 文献: Anti-melanogenic effects of epigallocatechin-3-gallate (EGCG), epicatechin-3-gallate (ECG) and gallocatechin-3-gallate (GCG) via down-regulation of cAMP/CREB /MITF signaling pathway in B16F10 melanoma cells
样品类型: Cell culture supernatant - 文献: rLj-RGD3, a novel recombinant toxin protein from Lampetra japonica, prevents coronary thrombosis-induced acute myocardial infarction by inhibiting platelet functions in rats
样品类型: platelets - 文献: Quinoline Derivative Enhances Human Sperm Motility and Improves the Functional Competence
样品类型: Spermatozoa (SPZ) - 文献: PKA-Dependent Membrane Surface Recruitment of CI-AMPARs Is Crucial for BCP-Mediated Protection Against Post-acute Ischemic Stroke Cognitive Impairment.
样品类型: Tissue homogenate - 文献: A novel role and mechanism of cystic fibrosis transmembrane conductance regulator in bisphenol A‐induced prostate cancer
样品类型: Cell culture supernatant - 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: Identification of a novel GNAS mutation in a case of pseudohypoparathyroidism type 1A with normocalcemia
样品类型: Cell culture supernatant - 文献: Estrogen Promotes cAMP Production in Mesenchymal Stem Cells by Regulating ADCY2
样品类型: Cell culture supernatant
