- 中文名称
RGM-A, ELISA BioAssay Kit (Repulsive Guidance Molecule A, RGM Domain Family Member A, Rgma)
- 英文名字
- RGM-A, ELISA BioAssay Kit (Repulsive Guidance Molecule A, RGM Domain Family Member A, Rgma)
- 供应商
- USBiological
- 产品货号
- R1994-87
- 产品报价
- ￥1/1Kit
- 产品说明书
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- 产品新闻
- 背景资料
- USBiological品牌是全球有名的抗原抗体和生化试剂供应商，生产世界上种类最多的抗体，用于Western Blot、免疫沉淀、免疫荧光、免疫组化和流式细胞术等多种检测方法。武汉艾美捷科技作为USBiological品牌中国区域总代理，是行业中少有的致力于服务客户，帮助客户，且拥有独立的专业销售团队、技术支持团队、市场营销团队、进出口报关团队的高科技生物企业。可以为您提供及时的咨询响应，专业的产品和解决方案支持，稳健快捷的交货周期，优质放心的售后服务。我们致力于为您提供有价值的产品和服务，在意您的成功！
- 产品描述
- Mouse Repulsive Guidance Molecule (RGM-A) is a 33kD member of the RGM family of GPI-linked membrane associated proteins (1, 2). It is synthesized as a 454aa prepro-protein that contains a 47aa N-terminal signal sequence and a 27aa C-terminal GPI attachment signal (3, 4). As a result of intramolecular cleavage, mature mouse RGM-A is a disulfide-linked dimer of the 122aa N-terminal and 258 C-terminal segments. The N-terminal segment contains an RGD tripeptide and two potential N-linked glycosylation sites, while the C-terminal segment contains an abbreviated von Willebrand factor domain (3-5). Mouse RGM-A shares 86%, 94%, and 100% aa sequence identity with chicken, human, and rat RGM-A, respectively. It also shares 56-48% aa sequence identity with RGM-B and -C, respectively.RGM-A is expressed in the developing central nervous system in a pattern that is complementary to the expression of RGM-B (3, 4). It is widely expressed on neuronal and glial cells of the mouse nervous system, particularly in the developing forebrain, the periventricular layers of the cerebrum, the optic tectum/superior colliculus of the midbrain, the retina, and in enteric ganglia of the fetal and adult gut (3-9). Among non-neuronal tissues, RGM-A mRNA is expressed in the gut, heart, lung, liver, skin, kidney, and testes (6, 8, 10).RGM-A regulates neuronal survival and development through interactions with a receptor complex that includes Neogenin and UNC5H2 (11, 12). The pattern of Neogenin expression overlaps tissue regions that exhibit RGM-A responsiveness (11, 13). RGM-A acts as a nervous system morphogen and as an axon guidance molecule. It promotes neuronal survival, inhibits neurite outgrowth, and induces growth cone collapse (5, 9, 12-15). RGM-A regulates the layer-specific termination of entorhinal complex axons into the hippocampus (16). It also guides retinal ganglion cell axons through the optic fissure to their accurate termination in the optic tectum (5, 17-20). Following tissue damage, RGM-A exerts prosurvival and neuroprotective effects on injured retinal ganglion cells (14). Its upregulation in the vicinity of spinal cord injuries can also hinder neurological regeneration by inhibiting axonal regeneration and synapse formation (7, 21).Similarly to other RGM family members, RGM-A also functions as a co-receptor that enhances signaling by BMP-2 and BMP-4 (10). The exact BMP signaling function that is mediated by RGM-A remains to be determined. Recently, RGM-B has been shown to be an important negative regulator of IL-6 expression in immune cells (22). The RGM-A gene has been shown to be associated with experimental inflammation and multiple sclerosis, suggesting that variants of the RGM-A gene can differentially affect immunoregulation and be associated with pro-inflammatory cytokines and antibody responses (23). In colorectal cancer, downregulation of RGM-A correlates with increased tumor cell proliferation and invasion (24).The Mouse/Rat RGM-A Immunoassay is a 4.5h solid phase ELISA designed to measure mouse and rat RGM-A in cell culture supernates, tissue homogenates, serum, and plasma. It contains NS0-expressed recombinant mouse RGM-A and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural RGM-A showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural RGM-A.Sample Type:Cell Culture Supernates-Remove particulates by centrifugation and assay immediately or aliquot and store samples at E -20°C. Avoid repeated freeze-thaw cycles.Tissue Homogenates-The preparation of tissue homogenates will vary depending upon tissue type. For this assay, brain tissue from individual mice was removed, rinsed in 1X PBS, and kept on ice. Using a tissue homogenizer, the tissue was homogenized in 1X PBS and frozen at £ -70°C. Cells were thawed and then frozen again for a total of two freeze-thaw cycles. After the second freeze-thaw cycle, the cells were centrifuged at 5000xg and the supernate was removed.Serum-Allow blood samples to clot for 2 hours at room temperature before centrifuging. Centrifuge for 20 minutes at 2000xg. Remove serum and assay immediately or aliquot and store samples at £ -20° C. Avoid repeated freeze-thaw cycles.Plasma-Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge for 20 minutes at 2000xg within 30 minutes of collection. Assay immediately or aliquot and store samples at £ -20° C. Avoid repeated freeze-thaw cycles.Note: Grossly hemolyzed samples are not suitable for use in this assay. Citrate plasma has not been validated for use in this assay.Sensitivity:Twenty-two assays were evaluated and the minimum detectable dose (MDD) of RGM-A ranged from 0.004-0.019ng/ml. The mean MDD was 0.010ng/ml. The MDD was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.Specificity:Recognizes both recombinant and natural mouse and rat RGM-A. The factors listed below were prepared at 500ng/m in Calibrator Diluent RD5-3 and assayed for cross-reactivity. Preparations of the following factors prepared at 500ng/ml in a mid-range RGM-A control were assayed for interference. No significant cross-reactivity or interference.Test Principle:This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for mouse RGM-A has been pre-coated onto a microplate. Standards, Control, and samples are pipetted into the wells and any RGM-A present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked polyclonal antibody specific for mouse RGM-A is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells. The enzyme reaction yields a blue product that turns yellow when the Stop Solution is added. The intensity of the color measured is in proportion to the amount of RGM-A bound in the initial step. The sample values are then read off the standard curve.Precision:Intra-assay:Mean (ng/ml): 0.190, 0.742, 2.12Standard Deviation: 0.010, 0.018, 0.087CV (%): 5.4, 2.4, 4.1Inter-assay:Mean (ng/ml): 0.195, 0.538, 2.07Standard Deviation: 0.015, 0.029, 0.147CV (%): 7.6, 5.3, 7.1Kit Components:Mouse/Rat RGM-A Microplate- 1x96 well polystyrene microplate (12 strips of 8 wells) coated with a monoclonal antibody specific for mouse RGM-A.Mouse/Rat RGM-A Conjugate- 1x12ml of a polyclonal antibody specific for mouse RGM-A conjugated to horseradish peroxidase with preservatives.Mouse/Rat RGM-A Standard-1x25ng of recombinant mouse RGM-A in a buffered protein base with preservatives; lyophilized.Mouse/Rat RGM-A Control- 1x1 vial of recombinant mouse RGM-A in a buffered protein base with preservatives; lyophilized. The concentration range of mouse RGM-A after reconstitution is shown on the vial label. The assay value of the Control should be within the range specified on the label.Assay Diluent RD1W- 1x12ml of a buffered protein solution with preservatives. Calibrator Diluent RD5-3-21ml of a buffered protein solution withpreservatives.Wash Buffer Concentrate- 50ml of a 25-fold concentrated solution of buffered surfactant with preservative.Color Reagent A 1x12m of stabilized hydrogen peroxide. Color Reagent B 1x12ml of stabilized chromogen (tetramethylbenzidine). Stop Solution- 1x23ml of diluted hydrochloric acid. Storage and Stability:Store components at 4°C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
- 产品特点
- 针对RGM-A, ELISA BioAssay Kit (Repulsive Guidance Molecule A, RGM Domain Family Member A, Rgma)该产品的特点优势，欢迎查阅官网提供的产品说明书。
- 保存建议
- 建议收到RGM-A, ELISA BioAssay Kit (Repulsive Guidance Molecule A, RGM Domain Family Member A, Rgma)产品后将其置于-20°C保存。
- 其他
- Usbiological公司是美国著名的抗体和生化试剂供应商，生产世界上种类最多的抗体，用于Western Blot、免疫沉淀、免疫荧光、免疫组化和流式细胞术等多种检测方法。Usbiological公司现已拥有超过50,19234种抗体、抗原和生化产品，为科研用户提供了诸多超值选择。
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- 注意
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