ELISA Kit for Fibroblast Growth Factor 9 (FGF9)
GAF; HBFG9; Glia-Activating Factor; Heparin-Binding Growth Factor 9
- Product No.SEA036Hu
- Organism SpeciesHomo sapiens (Human) 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 Length3h
- Detection Range15.6-1,000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 6.6pg/mL.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- DownloadInstruction Manual
- UOM48T96T96T*596T*1096T*100
- FOBUS$ 441 For more details, please contact local distributors!US$ 630 For more details, please contact local distributors!US$ 2835 For more details, please contact local distributors!US$ 5355 For more details, please contact local distributors!US$ 44100 For more details, please contact local distributors!
Packages (Simulation)- Packages (Simulation)
- Results demonstration
- Typical Standard Curve
- ISO9001: 2008, ISO13485: 2003 Registered
Specificity of the ELISA Kit for Fibroblast Growth Factor 9 (FGF9)
This assay has high sensitivity and excellent specificity for detection of Fibroblast Growth Factor 9 (FGF9).No significant cross-reactivity or interference between Fibroblast Growth Factor 9 (FGF9) and analogues was observed.
Recovery of the ELISA Kit for Fibroblast Growth Factor 9 (FGF9)
Matrices listed below were spiked with certain level of recombinant Fibroblast Growth Factor 9 (FGF9) and the recovery rates were calculated by comparing the measured value to the expected amount of Fibroblast Growth Factor 9 (FGF9) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 82-102 | 92 |
| EDTA plasma(n=5) | 91-101 | 98 |
| heparin plasma(n=5) | 90-98 | 94 |
Precision of the ELISA Kit for Fibroblast Growth Factor 9 (FGF9)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibroblast Growth Factor 9 (FGF9) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Fibroblast Growth Factor 9 (FGF9) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10%>Inter-Assay: CV<12%>
Linearity of the ELISA Kit for Fibroblast Growth Factor 9 (FGF9)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Fibroblast Growth Factor 9 (FGF9) 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) | 93-101% | 86-97% | 80-95% | 99-105% |
| EDTA plasma(n=5) | 83-95% | 79-101% | 84-102% | 83-93% |
| heparin plasma(n=5) | 82-101% | 79-95% | 84-101% | 78-98% |
Stability of the ELISA Kit for Fibroblast Growth Factor 9 (FGF9)
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 ELISA Kit for Fibroblast Growth Factor 9 (FGF9)
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 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;8. Add 50µL Stop Solution. Read at 450nm immediately.
Test principle of the ELISA Kit for Fibroblast Growth Factor 9 (FGF9)
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Fibroblast Growth Factor 9 (FGF9). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Fibroblast Growth Factor 9 (FGF9). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Fibroblast Growth Factor 9 (FGF9), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Fibroblast Growth Factor 9 (FGF9) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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Related products
| Catalog No. | Organism species: Homo sapiens (Human) | Applications (RESEARCH USE ONLY!) |
| RPA036Hu01 | Recombinant Fibroblast Growth Factor 9 (FGF9) | Positive Control; Immunogen; SDS-PAGE; WB. |
| APA036Hu01 | Active Fibroblast Growth Factor 9 (FGF9) | Cell culture; Activity Assays. |
| PAA036Hu01 | Polyclonal Antibody to Fibroblast Growth Factor 9 (FGF9) | WB; IHC; ICC; IP. |
| LAA036Hu71 | Biotin-Linked Polyclonal Antibody to Fibroblast Growth Factor 9 (FGF9) | WB; IHC; ICC. |
| MAA036Hu22 | Monoclonal Antibody to Fibroblast Growth Factor 9 (FGF9) | WB; IHC; ICC; IP. |
| SEA036Hu | ELISA Kit for Fibroblast Growth Factor 9 (FGF9) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| SCA036Hu | CLIA Kit for Fibroblast Growth Factor 9 (FGF9) | Chemiluminescent immunoassay for Antigen Detection. |
| PSA036Hu01 | Antibody Pair for Fibroblast Growth Factor 9 (FGF9) | ELISA,CLIA,ELISPOT,Luminex |
| KSA036Hu01 | ELISA Kit DIY Materials for Fibroblast Growth Factor 9 (FGF9) | Main materials for "Do It(ELISA Kit) Yourself" |
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另外还想问一下，还有其他可行的破碎方法吗
谢谢
问：1如才能提高柱子吸附蛋白量？
2样品中的核酸对吸附有无太大影响？
3我拟采用阴离子交换树脂，问pH8.0条件下除去核酸可否？
4怎么提高样品的澄清度？
以DNA凝胶电泳为例，电泳的操作步骤如下：
1、安装电泳槽
将有机玻璃的电泳凝胶床洗净，晾干，用胶带将两端的开口封好，放在水平的工作台上，插上样品梳。
2、琼脂糖凝胶的制备
称取琼脂糖溶解在电泳缓冲液中，（按0.3-1.5%的琼脂糖含量，1-25kb大小的DNA用1%的凝胶，20-100kb的DNA用0.5%的凝胶，200-2000bp的DNA用1.5%的凝胶）置微波炉或沸水浴中加热至完全溶化（不要加热至沸腾），取出摇匀。
3、灌胶
将冷却到60℃的琼脂糖溶液轻轻倒入电泳槽水平板上。
4、待琼脂糖胶凝固后，在电泳槽内加入电泳缓冲液，然后拔出梳子。
5、加样
将DNA样品（DNA样品是细胞破碎之后用离心管离心沉淀获得的）与加样缓冲液按4：1混匀后，用微量移液器将混合液加到样品槽中，每槽加10-20μl，记录样品的点样次序和加样量。
6、电泳
安装好电极导线，点样孔一端接负极，另一端接正极，打开电源，调电压至3-5V/cm，电泳1-3hr，当溴酚蓝移到距凝胶前沿1-2cm时，停止电泳。
7、染色和观察
取出凝胶，放在含有溴化乙锭的染色液中染色30min，即可在254nm的紫外灯下观察，有橙红色荧光条带的位置，即为DNA条带，或在紫外灯下照相记录电泳图谱。溴化乙锭是致癌剂，操作时要小心，必须戴手套。
扩展知识：
在外加直流电源的作用下，胶体微粒在分散介质里向阴极或阳极作定向移动，这种现象叫做电泳。利用电泳现象使物质分离，这种技术也叫做电泳。
凝胶电泳通常用于分析用途，但也可以作为制备技术，在采用某些方法（如质谱(MS)、聚合酶链式反应(PCR)、克隆技术、DNA测序或者免疫印迹）检测之前部分提纯分子。
琼脂糖 ( Agarose ) 是一种线性多糖聚合物 , 系从红色海藻产物琼脂中提取而来的。当琼脂糖溶液加热到沸点后冷却凝固便会形成良好的 电泳介 质 , 其密 度是由 琼脂 糖的浓 度决 定的。经过化学修饰的低熔点 (LMP) 的琼脂糖 , 在结构上比较脆弱 , 因此在较低的温度下便会熔化 , 可用于DNA片段的制备电泳。
聚丙烯酰胺凝胶主要有两种方式 : 一是用于分离和纯化双链DNA片段的非变性聚丙烯酰胺凝胶。在未变凝胶中分离 DNA的缺点是DNA的迁移 率受碱 基组成和序列的影响。由于无法得知未知DNA的迁移是否反常 , 故不能用未变性的聚丙烯酰胺凝胶电泳确定双链DNA的大小。二是用于分离及纯化单链DNA片 段的变性聚丙烯 酰胺凝 胶。这类聚丙烯酰胺凝胶是在核苷酸碱基配对抑制剂 ( 尿素或甲酰胺 ) 的存在下聚合而成 , 变性DNA的移动速度同其碱基组成及序列几乎完全无关 , 故可用于分离及纯化单链DNA片段和DNA测序等。
快速细胞破碎法鉴定转化子DNA（分离效果一般，二者要相差比较大才行，我常用前者，一般T载体上的要五百bp以上，一千bp以上最好！）
快速细胞破碎法鉴定转化子DNA
1、转化子菌落接种于相应的抗性培养基2.0ml，过夜培养。（要求设立对照！）
2、制备0.8%琼脂糖胶。
3、取1.5ml过夜培养物离心，15kr/min，1min；弃上清。（剩余保种！）
4、加水重悬，离心如上。留沉淀。
5、每管加50—100µl破碎细胞缓冲液(见后配方)，充分涡旋，使菌体悬浮均匀
6、水浴37℃，15min。
7、离心15kr/min，15min。
8、立即取20-30µl上清点样进行电泳，4—6V/cm。
9、电泳，当溴酚蓝到达凝胶板的2/3时，停止电泳。
10、染胶15—30min。
配方：50mMTris-HCl(pH6.8)；1%SDS；2mMEDTA；400mM蔗糖；0.01%溴酚兰
配法：1MTris-HCl(pH6.8),10ml;20%SDS,10ml;250mMEDTA,1.6ml;蔗糖27.2g;1.2%溴酚兰1.67ml加水至200ml;
煮沸法快速分析转化子DNA
1、挑单菌落于2.0ml相应抗生素液体培养基中，37℃振荡过夜（约16小时）。
2、取1.5ml过夜培养物于离心管中，15kr/min，1min。（剩余保种！）
3、弃上清，倒扣、流尽。
4、加入350µl由下列试剂组成溶液，混匀。
(8%蔗糖、0.5%TritonX-100、50mMEDTA(pH8.0)、10mMTris-HCl(pH8.0))
5、加入25µl新鲜配制的10mg/ml的溶菌酶（溶于10mMTris-HClpH8.0中），涡旋3--5sec。
6、立即将离心管浸入沸水浴40sec。
7、离心15kr/min，10min。
8、转移上清至另一离心管，加入2.5MNaAc40µl，加入异丙醇420µl，混匀后冰浴15min，沉淀DNA。
9、离心15kr/min，15min，将上层异丙醇弃去，真空抽干。
10、加入50µlTE缓冲液重悬，（含Rnase50µg/ml）。37℃保温10min。
11、取10µl样品点样，电泳，染胶。观察质粒DNA的区带位置。（要求有对照电泳！）
平常都是用超声破清洗器水浴超声破碎大约20分钟，然后上样的
但是一方面，常温超声不知道会不会造成蛋白降解破坏，因为发现加冰以后超声会变得非常弱，所以都么加冰。另一方面，因为水浴超声声强是不均匀的，如果样品多的话，会发现有些已经超碎DNA，但有些还是一团。
不知道是否有其他更好的方法？
