CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)
NTN; ABCD3; C3Xkine; CXC3; CXC3C; NTT; SCYD1; ABCD3; FKN; Neurotactin; Fractalkine; Small Inducible Cytokine Subfamily D(Cys-X3-Cys)Member 1
- Product No.SCA040Ra
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- All
- Human
- Mouse
- Rat
- Cavia
- Rabbit
- Simian
- Caprine
- Ovine
- Equine
- Bovine
- Porcine
- Gallus
- Canine
- Others
- Multi-species
- Pan-species
- Test MethodDouble-antibody Sandwich
- Assay Length2h, 40min
- Detection Range3.43-2,500pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 1.43pg/mL.
- Sample TypeSerum, plasma and other biological fluids
- DownloadInstruction Manual
- UOM48T96T96T*596T*1096T*100
- FOBUS$ 638 For more details, please contact local distributors!US$ 912 For more details, please contact local distributors!US$ 4104 For more details, please contact local distributors!US$ 7752 For more details, please contact local distributors!US$ 63840 For more details, please contact local distributors!
Specificity of the CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)
This assay has high sensitivity and excellent specificity for detection of Chemokine C-X3-C-Motif Ligand 1 (CX3CL1).No significant cross-reactivity or interference between Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) and analogues was observed.
Recovery of the CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)
Matrices listed below were spiked with certain level of recombinant Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) and the recovery rates were calculated by comparing the measured value to the expected amount of Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 95-102 | 98 |
| EDTA plasma(n=5) | 86-95 | 91 |
| heparin plasma(n=5) | 91-99 | 96 |
Precision of the CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10%>10%>Inter-Assay: CV<12%>12%>
Linearity of the CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 96-105% | 94-101% | 95-104% | 89-101% |
| EDTA plasma(n=5) | 78-103% | 78-102% | 97-104% | 80-97% |
| heparin plasma(n=5) | 83-101% | 78-103% | 80-93% | 88-102% |
Stability of the CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Assay procedure summary of the CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)
1. Prepare all reagents, samples and standards;2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;4. Aspirate and wash 3 times;5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;6. Aspirate and wash 5 times;7. Add 100µL Substrate Solution. Incubate 10 minutes at 37°C;8. Read RLU value immediately.
Test principle of the CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)
The microplate provided in this kit has been pre-coated with an antibody specific to Chemokine C-X3-C-Motif Ligand 1 (CX3CL1). Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Chemokine C-X3-C-Motif Ligand 1 (CX3CL1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) level in the sample or standard.;
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96-well Plate Assay Data Analysis Software
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Related products
| Catalog No. | Organism species: Rattus norvegicus (Rat) | Applications (RESEARCH USE ONLY!) |
| APA040Ra01 | Active Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) | Cell culture; Activity Assays. |
| RPA040Ra01 | Recombinant Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) | Positive Control; Immunogen; SDS-PAGE; WB. |
| PAA040Ra01 | Polyclonal Antibody to Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) | WB; IHC; ICC; IP. |
| LAA040Ra81 | FITC-Linked Polyclonal Antibody to Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) | WB; IHC; ICC; IF. |
| LAA040Ra71 | Biotin-Linked Polyclonal Antibody to Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) | WB; IHC; ICC. |
| MAA040Ra21 | Monoclonal Antibody to Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) | WB; IHC; ICC; IP. |
| SEA040Ra | ELISA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| SCA040Ra | CLIA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) | Chemiluminescent immunoassay for Antigen Detection. |
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在样品用液氮瞬间冻结之后,应该储存在-80°C,千万不能解冻。即使是置于含有胍盐的裂解液中作匀浆前的短暂解冻,也会导致RNA的降解和损失。瞬间冻结的组织应该首先在超低温条件下先研磨成粉,然后置于裂解液中进行匀浆。
RNAfixer使样品储存更为便利。储存在RNAfixer中的细胞或组织可在室温下稳定保存长达1个星期,在4°C可稳定保存长达1个月,或永久保存在-20°C。
2. 消除环境RNase的污染
为了得到完整的、高品质RNA,在整个RNA制备过程中,当RNA离开强蛋白变性剂(如离液裂解液或酚)的保护时,避免引入新的RNase污染就非常关键。由于RNase几乎是无所不在,所以必须确保与纯化的RNA接触的每一样东西都是无RNase污染的。所有的表面,包括移液器、工作台、玻璃器皿和制胶设备,都必须用表面去污净化溶液如RNase喷雾清除剂 来处理过,去除各种溶液或者反应缓冲液中可能存在的RNase污染,可以用RNAsafe。必须保证一直使用无RNase的枪头、试管和溶液,手套也应经常更换。
3.迅速灭活内源的RNA酶,以防止RNA降解。
以下3个方法均可有效使内源RNA酶失活:
1)、用含离液(如胍盐)的细胞裂解液收获样品,并立即匀浆。
2)、用液氮瞬间冻结样品。值得特别注意的是:组织块必须保证足够小,在浸入液氮的瞬间就能冻结,以确保瞬间令RNA酶失活。
3)、立即将样品置于RNAfixer无液氮RNA样品储存液中。它是一种水相、无毒的收集试剂,能立即稳定并保护完整、未冻结的组织和细胞样品中的RNA。关键要点是组织样品切片一定要够薄(<0.5 cm),这样RNAfixer才能在RNase破坏RNA之前迅速渗入组织块中。
4. 选择合适破壁方法
细胞或组织的彻底匀浆对RNA提取来说,是一个很关键的步骤,它能够防止RNA的损失和降解。匀浆的方法应根据细胞或组织的类型来选择。大部分培养的细胞可以置于细胞裂解液中,通过简单的涡旋震荡来匀浆;而动物组织、植物组织、酵母和细菌、真菌则常常需要更加剧烈的方法,通常用液氮研磨。比如说细菌(特别是革兰氏阳性菌)的细胞壁,就需要溶菌酶消化来实现彻底的细胞裂解和RNA的最大回收,酵母提取时加入破壁酶帮助破壁,再用TRIpure或RNApure进行提取。
5. RNA提取少走弯路——不同材料如何选择最适RNA提取试剂
现有众多的RNA分离方法也许令人难以取舍。目前最简单也是最安全的方法是柱式分离,如RNApure 因为操作简单,时间短,纯度高而受到大家的喜爱,RNApure不需要DNA酶消化DNA,节省了时间,避免RNA的降解,从而提高了产量;相对非柱式分离(如TRIzol),去除蛋白及其他杂质干净,提高了纯度,对于细胞、组织、一般植物均非常适合。植物RNA的提取比较难抉择,植物RNA提取受酚、多糖、蛋白杂质、次生代谢物含量的影响,一般用TRIzol提取纯度不高,而且很多植物用TRIzol提取不出来。
这时候可以选择多糖多酚植物总RNA提取试剂盒(水仙、辣椒、胡萝卜、玉米、百合、小麦、西红柿、花菜、油菜等)和通用植物RNA提取试剂盒(适合绝大多数植物提取,包括:各种中草药、植物种子、苹果、葡萄、草莓、香蕉、龙眼、荔枝、草坪植物、松树、杉树、白桦、紫松果、彩叶草、一品红、夹竹桃、垂叶榕、紫罗兰、月季、天竺蓝、牵牛花等);非常值得一提的是血液(包括血清、血浆、脑脊液、各种拭子或其他液体含量高的样本)RNA的提取用TRIzol和红细胞裂解的方法效果都不好,因为红细胞裂解液不含有RNA酶的抑制成分,这个过程RNA很容易降解,推荐使用TRIpure LS (RP1101)或者血液总RNA提取试剂盒(RP4001),该方法适用于表达谱芯片实验中RNA的提取。
6. 低浓度RNA的沉淀
纯化得到的RNA可能需要通过沉淀来浓缩,以满足一些下游应用的需要。醋酸铵(NH4OAc) 沉淀(加0.1体积的5M 醋酸铵、2-2.5体积的无水乙醇,-20°C放置25分钟以上)可以很好地回收RNA。如果需要定量回收低浓度的RNA(ng/ml),可以采用共沉淀(如linear acrylamide糖原glycogen,、酵母yeast RNA )的方法。核酸助沉剂是linear acrylamide,当RNA用于RT-PCR分析时,线性的丙烯酰胺和DNase处理的糖原都可以作为理想的共沉淀剂,因为它们都不含DNA污染,糖原含量高会抑制PCR反应,应注意控制浓度,核酸助沉剂对PCR无影响,成为病毒核酸提取的首选。酵母RNA和未处理的糖原会给样品带来核酸污染,有可能影响RT-PCR的结果。沉淀后,注意避免RNA沉淀过分干燥,因为这可能导致很难重新溶解。
7. 提取好的RNA如何储存
如果只是短期储存,重悬的RNA应放置于-20°C;如果是长期储存的话,就应该放置于-80°C。储存RNA时,可以加入少量的RNA酶抑制剂(RP5601),避免RNA的降解,RNA可以直接做下游实验。如果要长期保存RNA,可以加入RNAlong。我们推荐将RNA溶液分装在几支管中。这会避免反复冻融损伤RNA,并预防偶然的RNase污染。
氯仿萃取RNA
异丙醇沉淀RNA
酒精清洗
Trizol法和试剂盒提取组织中的RNA,哪个方法更可靠和精确呢?谢谢!
1.破碎和裂解植物细胞壁。
2.抑制rna酶
3.去除植物多糖和多酚。树脂 淀粉和纤维材料。
4.去除许多高等植物组织尤其是成熟组织能产生某些水溶性的次级代谢产物,这些次级代谢产物很容易与RNA结合并与RNA共同被抽提出来而阻碍具有生物活性的RNA分离。
除了干扰RNA分离之外,植物多糖和多酚及次级代谢产物还能明显抑制下游RNA反应如转录和PCR.
1、RNA提取为什么用异丙醇,而且是等体积的?
2、为什么用氯仿,RNA为什么在上清中,中间和下层分别有什么?
3、RNA提取最后为什么用75%乙醇洗,而且为什么使用的是75%乙醇而不用无水乙醇来洗。还有有人建议用完75%乙醇后用无水乙醇洗,会使水分快速蒸发,可行吗?
谢谢大家啦,帮帮忙。
RNAi技术的原理与应用(一).ppt(218.5k)
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