1.从少量样品(1-10mg组织或102-104细胞)中提取RNA时可加入少许糖原以促进RNA沉淀。例如加800ml TRIzol匀浆样品,沉淀RNA前加5-10μg RNase-free糖原。糖原会与RNA一同沉淀出来,糖原浓度不高于4mg/ml是不影响一链的合成,也不影响PCR反应。
1. 匀浆处理:a.组织 将组织在液氮中磨碎,每50-100mg组织加入1ml TRIzol,用匀浆仪进行匀浆处理。样品体积不应超过TRIzol体积10℅。
b.单层培养细胞 直接在培养板中加入TRIzol裂解细胞,每10cm2面积(即3.5cm直径的培养板)加1ml,用移液器吸打几次。TRIzol的用量应根据培养板面积而定,不取决于细胞数。TRIzol加量不足可能导致提取的RNA有DNA污染。
c.细胞悬液 离心收集细胞,每5-10×106动物、植物、酵母细胞或1×107细菌细胞加入1ml TRIzol,反复吸打。加TRIzol之前不要洗涤细胞以免mRNA降解。一些酵母和细菌细胞需用匀浆仪处理。
3.可选步骤:如样品中含有较多蛋白质,脂肪,多糖或胞外物质(肌肉,植物结节部分等)可于2-8℃10000×g离心10分钟,取上清。离心得到的沉淀中包括细胞外膜,多糖,高分子量DNA,上清中含有RNA。处理脂肪组织时,上层有大量油脂应去除。取澄清的匀浆液进行下一步操作。
5. 每使用1ml TRIzol加入0.2ml,剧烈振荡15秒,室温放置3分钟。
6. 2-8℃10000×g离心15分钟。样品分为三层:底层为黄色有机相,上层为无色水相和一个中间层。RNA主要在水相中,水相体积约为所用TRIzol试剂的60℅。
7. 把水相转移到新管中,如要分离DNA和蛋白质可保留有机相,进一步操作见后。用异丙醇沉淀水相中的RNA。每使用1ml TRIzol加入0.5ml异丙醇,室温放置10分钟。
8. 2-8℃10000×g离心10分钟,离心前看不出RNA沉淀,离心后在管侧和管底出现胶状沉淀。移去上清。
9. 用75℅乙醇洗涤RNA沉淀。每使用1ml TRIzol至少加1ml 75℅乙醇。2-8℃不超过7500×g离心5分钟,弃上清。
10.室温放置干燥或真空抽干RNA沉淀,大约晾5-10分钟即可.不要真空离心干燥,过于干燥会导致RNA的溶解性大大降低。加入25-200μl无RNase的水或0.5℅SDS,用枪头吸打几次,55-60℃放置10分钟使RNA溶解.如RNA用于酶切反应,勿使用SDS溶液。RNA也可用100℅的去离子甲酰胺溶解,-70℃保存。
研究领域 肿瘤 细胞生物 发育生物学
蛋白分子量 predicted molecular weight: 27kDa
性 状 Lyophilized or Liquid
免 疫 原 KLH co
njugated synthetic peptide derived from human C4orf22
亚 型 IgG
纯化方法 affinity purified by Protein A
储 存 液 Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4
产品应用 WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 ICC=1:100-500 IF=1:100-500
(石蜡切片需做抗原修复)
not yet tested in other applications.
optimal dilutions/co
ncentrations should be determined by the end user.
保存条件 Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one mo
nth and for greater than a year when kept at -20°C. When reco
nstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.
im
portant Note This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
产品介绍 C4orf22 is a 233 amino acid protein that exists as three alternatively spliced isoforms and are encoded by a gene encoding maping to human chromosome 4, which represents approximately 6% of the human genome and co
ntains nearly 900 genes. Notably, the Huntingtin gene, which is found to encode an expanded glutamine tract in cases of Huntington's disease, is on chromosome 4. FGFR-3 is also encoded by a gene that maps to chromosome 4 and has been associated with thanatophoric dwarfism, achondro
plasia, Muenke syndrome and bladder cancer. Chromosome 4 is also tied to Ellis-van Creveld syndrome, methylmalo
nic acidemia and polycystic kidney disease.
Databa
se l
inks : UniProtKB/Swiss-Prot: Q6V702.4
英文名称 Anti-C4orf14
中文名称 4号染色体开放阅读框14抗体
别 名 mAtNOS1; Chromosome 4 open reading f
rame 14; hAtNOS1; Hypothetical protein LOC84273; MGC3232; Putative ortholog of Arabidopsis mitocho
ndrial nitric oxide synthase; Uncharacterized protein C4orf14; NOA1_HUMAN.
浓 度 1mg/1ml
规 格 0.2ml/200μg
抗体来源 Rabbit
克隆类型 polyclonal
交叉反应 Human, Mouse, Rat
产品类型 一抗
研究领域 肿瘤 细胞生物 免疫学 发育生物学
蛋白分子量 predicted molecular weight: 78kDa
性 状 Lyophilized or Liquid
免 疫 原 KLH co
njugated synthetic peptide derived from human C4orf14
亚 型 IgG
纯化方法 affinity purified by Protein A
储 存 液 Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4
产品应用 WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 ICC=1:100-500 IF=1:100-500
(石蜡切片需做抗原修复)
not yet tested in other applications.
optimal dilutions/co
ncentrations should be determined by the end user.
保存条件 Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one mo
nth and for greater than a year when kept at -20°C. When reco
nstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.
im
portant Note This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
产品介绍 Representing approximately 6% of the human genome, chromosome 4 co
ntains nearly 900 genes. Notably, the Huntingtin gene, which is found to encode an expanded glutamine tract in cases of Huntington's disease, is on chromosome 4. FGFR-3 is also encoded on chromosome 4 and has been associated with thanatophoric dwarfism, achondroplasia, Muenke syndrome and bladder cancer. Chromosome 4 is also tied to Ellis-van Creveld syndrome, methylmalo
nic acidemia and polycystic kidney disease. Chromosome 4 reportedly co
ntains the largest gene deserts (regions of the genome with no protein encoding genes) and has one of the two lowest recombination frequencies of the human chromosomes. The C4orf14 gene product has been provisio
nally designated C4orf14 pending further characterization.
固相内毒素清除剂100g价格Function : Involved in regulation of mitocho
ndrial protein translation and respiration. Plays a role in mitochondria-mediated cell death. May act as a scaffolding protein or stabilizer of respiratory chain supercomplexes. Binds GTP.
Subunit : Homodimer or multimer (By similarity). Interacts with mitocho
ndrial complex I, DAP3, MRPL12 and MRPS27.
Subcellular Location : Mitocho
ndrion inner membrane; Peripheral membrane protein; Matrix side.