Product Details
- Target Species
- Bovine
- Species Cross-Reactivity
Target Species Cross Reactivity Horse Sheep - N.B. Antibody reactivity and working conditions may vary between species.
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
0.09% Sodium Azide - Carrier Free
- Yes
- Immunogen
- Plasmid cDNA encoding bovine IL-10.
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Fusion Partners
- Spleen cells from immunised Balb/c mice were fused with cells of the mouse SP2/0 myeloma cell line
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred. This product should be stored undiluted.Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
- Guarantee
- 12 months from date of despatch
More Information
- UniProt
- P43480
- Entrez Gene
- IL10
- GO Terms
- GO:0002237response to molecule of bacterial origin
- GO:0002740negative regulation of cytokine secretion involved in immune response
- GO:0002904positive regulation of B cell apoptosis
- GO:0005125cytokine activity
- GO:0005615extracellular space
- GO:0006954inflammatory response
- GO:0006955immune response
- GO:0030889negative regulation of B cell proliferation
- GO:0032715negative regulation of interleukin-6 production
- GO:0032800receptor biosynthetic process
- GO:0042095interferon-gamma biosynthetic process
- GO:0043193positive regulation of gene-specific transcription
- GO:0045019negative regulation of nitric oxide biosynthetic process
- GO:0045077negative regulation of interferon-gamma biosynthetic process
- GO:0050715positive regulation of cytokine secretion
- GO:0051045negative regulation of membrane protein ectodomain proteolysis
- GO:0051091positive regulation of transcription factor activity
- GO:0051384response to glucocorticoid stimulus
- Regulatory
- For research purposes only
Applications of IL-10 antibody
Application Name | Verified | Min Dilution | Max Dilution |
---|---|---|---|
ELISA | 5ug/ml | 10ug/ml | |
ELISpot | |||
Flow Cytometry 1 |
- 1 Membrane permeabilization is required for this application. Bio-Rad recommends the use of Leucoperm (BUF09) for this purpose.
- ELISA
- This reagent may be used as a capture antibody in a sandwich ELISA assay for bovine IL-10 in combination with MCA2111B as detection reagent, see Bannerman, D.D.et al.
Secondary Antibodies Available
Description | Product Code | Applications | Pack Size | List Price | Quantity |
---|---|---|---|---|---|
Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed) | STAR117A | E WB | 0.5 mg | ||
Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed) | STAR117F | F | 0.5 mg | ||
Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed) | STAR117P | E WB | 0.5 mg | ||
Goat anti Mouse IgG (Fc):FITC | STAR120F | C F | 1 mg | ||
Goat anti Mouse IgG (Fc):HRP | STAR120P | E WB | 1 mg | ||
Rabbit F(ab')2 anti Mouse IgG:RPE | STAR12A | F | 1 ml | ||
Goat anti Mouse IgG:FITC (Rat Adsorbed) | STAR70 | F | 0.5 mg | ||
Goat anti Mouse IgG:RPE (Rat Adsorbed) | STAR76 | F | 1 ml | ||
Goat anti Mouse IgG:HRP (Rat Adsorbed) | STAR77 | C E P | 0.5 mg | ||
Goat anti Mouse IgG/A/M:Alk. Phos. | STAR87A | C E WB | 1 mg | ||
Goat anti Mouse IgG/A/M:HRP (Human Adsorbed) | STAR87P | E | 1 mg | ||
Rabbit F(ab')2 anti Mouse IgG:Dylight®800 | STAR8D800GA | F IF WB | 0.1 mg | ||
Rabbit F(ab')2 anti Mouse IgG:FITC | STAR9B | F | 1 mg | ||
Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed) | STAR13B | C E P RE WB | 1 mg | ||
Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed) | STAR117D800GA | F IF WB | 0.1 mg | ||
Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed) | STAR117D488GA | F IF | 0.1 mg | ||
Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed) | STAR117D680GA | F WB | 0.1 mg |
Negative Isotype Controls Available
Description | Product Code | Applications | Pack Size | List Price | Quantity |
---|---|---|---|---|---|
Mouse IgG2b Negative Control | MCA691 | F | 100 Tests |
Application Based External Images
ELISA
ELISpot
Flow Cytometry
Product Specific References
References for IL-10 antibody
- Kwong, L.S. et al. (2002) Development of an ELISA for bovine IL-10.Vet Immunol Immunopathol. 85 (3-4): 213-23.
- Scandurra, G.M. et al. (2009) Assessment of live candidate vaccines for paratuberculosis in animal models and macrophages.Infect Immun. 78: 1383-9.
- Weiss DJ et al. (2008) Bovine monocyte TLR2 receptors differentially regulate the intracellular fate of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium.J Leukoc Biol. 83 (1): 48-55.
- Hamza, E. et al. (2007) Modulation of allergy incidence in icelandic horses is associated with a change in IL-4-producing T cells.Int Arch Allergy Immunol. 144: 325-37.
- Wenz, J.R. et al. (2010) Factors associated with concentrations of select cytokine and acute phase proteins in dairy cows with naturally occurring clinical mastitis.J Dairy Sci. 93: 2458-70.
- Rinaldi, M. et al (2010) A sentinel function for teat tissues in dairy cows: dominant innate immune response elements define early response to E. coli mastitis.Funct Integr Genomics. 10: 21-38.
- Parker, D.G. et al. (2010) Lentivirus-mediated gene transfer of interleukin 10 to the ovine and human cornea.Clin Experiment Ophthalmol. 38: 405-13.
- Ferret-Bernard, S. et al. (2011) Mesenteric lymph node cells from neonates present a prominent IL-12 response to CpG oligodeoxynucleotide via an IL-15 feedback loop of amplification.Vet Res. 42:19.
- Bannerman, D.D. et al. (2004) Escherichia coli and Staphylococcus aureus elicit differential innate immune responses following intramammary infection.Clin Diagn Lab Immunol. 11: 463-72.
- Coad, M. et al. (2010) Repeat tuberculin skin testing leads to desensitisation in naturally infected tuberculous cattle which is associated with elevated interleukin-10 and decreased interleukin-1 beta responses.Vet Res. 41: 14.
- den Hartog, G. et al. (2011) Modulation of human immune responses by bovine interleukin-10.PLoSone 6: e18188
- Ikebuchi, R. et al. (2013) Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro.Vet Res. 44: 59.
- Ferret-Bernard, S. et al. (2010) Cellular and molecular mechanisms underlying the strong neonatal IL-12 response of lamb mesenteric lymph node cells to R-848.PLoS One. 5: e13705.
- Jones, G.J. et al. (2010) Simultaneous measurement of antigen-stimulated interleukin-1 beta and gamma interferon production enhances test sensitivity for the detection of Mycobacterium bovis infection in cattle.Clin Vaccine Immunol. 17: 1946-51.
- McGill, J.L. et al. (2013) Differential chemokine and cytokine production by neonatal bovine γ/δ T-cell subsets in response to viral toll-like receptor agonists and in vivo respiratory syncytial virus infection.Immunology. 139: 227-44.
- Olivier, M. et al. (2009) Capacities of migrating CD1b+ lymph dendritic cells to present Salmonella antigens to naive T cells.PLoS One. 7: e30430.
- Shu, D. et al. (2011) Diverse cytokine profile from mesenteric lymph node cells of cull cows severely affected with Johne"s disease.Clin Vaccine Immunol. 18: 1467-76.
- Redondo, E. et al. (2014) Induction of interleukin-8 and interleukin-12 in neonatal ovine lung following experimental inoculation of bovine respiratory syncytial virus.J Comp Pathol. 150 (4): 434-48.
- Dooley LM et al. (2015) Effect of mesenchymal precursor cells on the systemic inflammatory response and endothelial dysfunction in an ovine model of collagen-induced arthritis.PLoS One. 10 (5): e0124144.
- Rainard, P. et al. (2016) Innate and Adaptive Immunity Synergize to Trigger Inflammation in the Mammary Gland.PLoS One. 11 (4): e0154172.
- Canal AM et al. (2017) Immunohistochemical detection of pro-inflammatory and anti-inflammatory cytokines in granulomas in cattle with natural Mycobacterium bovis infection. Res Vet Sci. 110: 34-39.
- Cassady-Cain, R.L. et al. (2017) Inhibition of Antigen-Specific and Nonspecific Stimulation of Bovine T and B Cells by Lymphostatin from Attaching and Effacing Escherichia coli.Infect Immun. 85 (2)Jan 26 [Epub ahead of print].
- Pomeroy B et al. (2016) Impact of in vitro treatments of physiological levels of estradiol and progesterone observed in pregnancy on bovine monocyte-derived dendritic cell differentiation and maturation.Vet Immunol Immunopathol. 182: 37-42.
- Davidson, J.O. et al. (2020) Window of opportunity for human amnion epithelial stem cells to attenuate astrogliosis after umbilical cord occlusion in preterm fetal sheep.Stem Cells Transl Med. Oct 26 [Epub ahead of print].
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白细胞介素10(IL-10)是一种由多种细胞亚群产生的重要免疫调节因子。
早期研究显示,其主要生物功能为限制炎症反应及调节多种免疫细胞的分化和增殖,例如:T细胞、B细胞、NK细胞、
抗原呈递细胞、肥大细胞、粒细胞等。近来研究结果提示,IL-10也可以介导免疫反应的激活,有助于清除感染及非感染
颗粒且只产生轻微的炎症反应。大量的研究,包括患者IL-10的表达分析、体外实验及动物实验表明,IL-10与自身免疫
性疾病、炎症、肿瘤等疾病的发生、发展关系密切。临床试验主要集中于RA、炎症性肠病、银屑病、器官移植和丙型肝
炎等。虽然试验结果各不相同,但是使我们对IL-10在免疫反应中的作用的认识更加深入,同时也预示IL-10可以成为一
种治疗药物。目前,我国还没有重组基因工程IL-10药物上市,而且在探索如何高效表达具有生物活性rhIL-10的研究尚
处于起步阶段。本研究建立在国内外IL-10重组基因工程研究基础上,拟克隆人IL-10基因,通过原核、真核两种表达系
统表达IL-10蛋白,奠定重组基因工程研制人IL-10的基础。主要研究结果如下:一、依据Genbank中人类IL-10序列设计
一对特异性引物,应用RT-PCR技术,从LPS刺激的外周血单个核细胞中成功地克隆出hIL-10成熟肽编码序列,并将之克
隆入pGEM-T载体,构建出中间载体<WP=91>pGEM-T-hIL-10,转化入大肠杆菌JM109中,经酶切鉴定及序列测定与
分析,与GenBank中发表的(NM000572.2)hIL-10成熟肽编码序列完全一致。为hIL-10基因的表达研究奠定了基
础。二、从中间载体pGEM-T-hIL-10中将hIL-10成熟肽编码序列酶切回收,定向克隆至原核表达载体pET-28a(+)的T7
启动子下游,成功地构建了原核表达载体pET-28a(+)-hIL-10,将表达载体转化入大肠杆菌BL21(DE3)pLyss中获得表达
菌株,经IPTG的诱导获得较高表达,表达蛋白以包涵体形式存在。经薄层扫描确定目的蛋白表达量占菌体总蛋白的
28.7%。Westernblotting分析结果显示,表达产物与抗hIL-10单克隆抗体呈特异性阳性反应。产物经固相金属离子亲
和层析方法初步纯化,并在柱上应用浓度递减的尿素缓冲液对产物进行复性。活性研究结果表明,复性产物具有抑制
LPS刺激的外周血单核细胞表达MHCⅡ分子的活性。三、为了进一步获得高活性、高表达、适用于产业化生产的rhIL-
10,本研究选择应用酵母表达系统对hIL-10进行表达。应用PCR方法从pGEM-T-hIL-10扩增出hIL-10成熟肽编码序
列,并在其上下游分别引入EcoRI和BaxⅠ两个酶切位点,通过定向克隆成功地构建出分泌型酵母表达载体pPICZαA-
rhIL-10。采用电穿孔技术将表达单位转化入酵母菌P.pastorisX-33中,通过提取酵母基因组进行PCR鉴定以及
MD/MM营养表型筛选,初步筛选出10株可表达具有hIL-10活性的酵母菌。小量发酵培养表达菌株,以0.5%甲醇诱导
96小时,培养上清中蛋白经硫酸铵沉淀后,经SDS-PAGE、薄层扫描显示rhIL-10约占样品蛋白总量的10%。产物经固
相金属离子亲和层析方法初步纯化,纯度达94%。活性研究结果显示酵母表达的分泌型rhIL-10具有较原核表达产物更高
的活性。
盐沉淀
Epicentre(Illumina旗公司)MasterPure™ Complete DNA Purification Kit采用种专利盐沉淀系列试剂盒覆盖各种品类型包括血液、固定组织、植物、酵母、昆虫等短短30钟内带高产量核酸损失极少
据Epicentre资深产品经理Cristine Kinross介绍些试剂盒用户测序或其物应用品制备前需再进行额外纯化除蛋白及其细胞碎片引入些必须除毒性化合物避免洗涤并改善核酸产量
Kinross谈道:利用MasterPure收核酸色纯度让品直接进入测序文库制备于其应用包括芯片、qPCR克隆适合外试剂盒能用于RNA纯化获总核酸经DNase处理能总RNA
种试剂盒
Promega提供广泛核酸纯化试剂盒采用几种同策略例Wizard® Genomic DNA Purification Kit基于经典沉淀全血、组织培养细胞、物及植物组织、酵母细菌提取基组DNA灵高效;ReliaPrep™ 系列试剂盒采用柱提简单快速外Promega提供基于磁珠纯化试剂盒
Promega公司基组全球产品经理Eric Vincent表示:尽管溶液确切组同裂解、结合、洗涤洗脱基本程适用于同结合(硅胶或纤维素)同纯化形式(纯化柱或磁珠)及手自纯化操作于些非重要品类型(FFPE品)需要些专门操作或处理
我发独特结合/洗涤缓冲液能效除抑制游反应物质优化系统让研究员能够捕获少量核酸并体积(30 μl )洗脱我发自化解决案适合各种试剂通量Vincent说
些试剂盒提供完整解决案能够极体积洗脱核酸同避免抑制剂残留洗脱液经些试剂盒纯化核酸用于PCR、Sanger测序、新代测序、芯片等游析
经典硅胶膜
QIAGEN核酸纯化试剂盒依赖DNA与纯化柱硅胶膜结合抑制剂洗掉完整DNA随柱洗脱产量高纯度高带更高质量新代测序文库
Marco Polidori - QIAGEN本制备全球产品经理 - 认处理极少量品或困难品FFPE高产量变格外关键FFPE品所产DNA能现随机胞嘧啶脱氨事件导致单核苷酸态性(SNP)QIAGENGeneRead DNA FFPE Kit能够除脱氨胞嘧啶避免NGS实验产错误结产品尚未式推若兴趣试用填写资料
我部试剂盒都已NGS应用证明循环肿瘤DNA全基组测序微物组Polidori谈道些试剂盒能让用户各种品获高质量DNA避免FFPE品现C-T假象并短间内带高量DNAMagAttract HMW DNA kit能够利用简单磁珠操作实现100-200 kb DNA纯化整纯化程温除污染物及抑制剂并具极高产量纯度
离液盐
Sigma-Aldrich公司提供GenElute™ Mammalian Genomic DNA Purification Kit基于离液盐试剂产品含离液盐溶液品裂解确保彻底变性添加乙醇让DNA与硅胶膜结合污染物洗掉DNA洗脱Tris缓冲液
通专离基组DNA选择硅胶膜试剂盒让用户能够各种品纯化高质量DNA包括培养细胞、组织(包括鼠尾)、新鲜全血或白细胞
据介绍试剂盒综合硅胶膜结合微量离形式处避免昂贵树脂、乙醇沉淀及害机物苯酚、氯仿更重要纯化所DNA应用于限制性内切酶消化、PCR、Southern blot、测序等
(作者:James Netterwald/物通编译)
第十二条物制品产环境空气洁净度级别应与产品产操作相适应厂房与设施应原料、间体品造污染
第十三条产程涉及高危操作其空气净化系统等设施应符合特殊要求
第十四条物制品产操作应符合表规定相应级别洁净区内进行未列操作参照表适级别洁净区内进行:
洁净度级别
物制品产操作示例
B级背景局部A级
附录菌药品非终灭菌产品规定各工序
灌装前经除菌滤制品其配制、合并等
C级
体外免疫诊断试剂阳性血清装、抗原与抗体装
D级
原料血浆合并、组离、装前巴氏消毒
口服制剂其发酵培养密闭系统环境(暴露部需菌操作)
酶联免疫吸附试剂等体外免疫试剂配液、装、干燥、内包装
第十五条产程使用某些特定物体阶段应根据产品特性设备情况采取相应预防交叉污染措施使用专用厂房设备、阶段性产式、使用密闭系统等
第十六条灭疫苗(包括基重组疫苗)、类毒素细菌提取物等产品灭交替使用同灌装间灌装、冻干设施每装应采取充污染措施必要应进行灭菌清洗
第十七条卡介苗结核菌素产厂房必须与其制品产厂房严格产涉及物产设备应专用
第十八条致病性芽胞菌操作直至灭程完前应使用专用设施炭疽杆菌、肉毒梭状芽胞杆菌破伤风梭状芽胞杆菌制品须相应专用设施内产
第十九条其种类芽孢菌产品某设施或套设施期轮换产芽胞菌制品任何间能产种产品
第二十条使用密闭系统进行物发酵同区域同产单克隆抗体重组DNA制品
第二十条菌制剂产加工区域应符合洁净度级别要求并保持相压;操作致病作用微物应专门区域内进行并保持相负压;采用菌工艺处理病原体负压区或物安全柜其周围环境应相压洁净区
第二十二条菌(毒)操作区应独立空气净化系统自病原体操作区空气循环使用;自危险度二类病原体操作区空气应通除菌滤器排放滤器性能应定期检查
第二十三条用于加工处理物体产操作区设备应便于清洁污染清洁污染效性应经验证
第二十四条用于物体培养设备应能够防止培养物受外源污染
第二十五条管道系统、阀门呼吸滤器应便于清洁灭菌宜采用线清洁、线灭菌系统密闭容器(发酵罐)阀门应能用蒸汽灭菌呼吸滤器应疏水性材质且使用效期应经验证
第二十六条应定期确认涉及菌毒种或产品直接暴露隔离、封闭系统泄漏风险
第二十七条产程病原体污染物品设备应与未使用灭菌物品设备并明显标志
第二十八条产程需要称量某些添加剂或(缓冲液)产区域存放少量物料
第二十九条洁净区内设置冷库恒温室应采取效隔离防止污染措施避免产区造污染
面GMP附录3要求关键看体外试剂哪类般没问题
1.相同房间产段产
2.同房间产且使用同空调系统
我非医学专业人士,但因工作需要了解这家公司和它的产品品质,希望用过或对比过的童鞋能帮我科普下专业知识,万分感谢~~具体问题可以私聊哈,如果能留下QQ、电话最佳~~~
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