AmplideX® PCR/CE FMR1 Reagents
AmplideX PCR/CE FMR1 Reagents* are market-leading research tools for the detection of CGG repeats in the fragile X mental retardation (FMR1) gene. These reagents provide a PCR-only approach based on Triplet Repeat Primed PCR (TP-PCR) design to reliably amplify and detect all alleles including Full Mutations.
Features & Benefits
AmplideX PCR/CE FMR1 Reagents* have created an easy-to-use, accessible, high performance method for laboratories to reliably analyze CGG repeats and detect interrupting AGG sequences in the FMR1 gene.
Reduced ComplexityEase-of-analysis of the FMR1 gene has been simplified through:
- Implementation of proprietary PCR solution for amplifying GC-rich regions
- Automation of result calling using AmplideX PCR/CE FMR1 Reporter*
Optimized WorkflowValuable operator hands-on time has been significantly reduced through:
- Direct injection of PCR products (no PCR clean up) in to Capillary Electrophoresis platforms
- Decreased need for Southern blot analysis (up to 50 fold)
- End-to-end solution for FMR1 analysis including all necessary reagents and software
Quality PerformancePerforming FMR1 Analysis with Greater Sensitivity and Accuracy:
- Detection of all allele expansions, including low abundance full mutation size mosaics with up to at least 1300 CGG repeats
- Up to 875 fold more sensitive than Southern blot1
- Resolution of female homozygous and heterozygous samples and indication of interrupting AGG sequences
- Proven performance as indicated by more than 30 peer reviewed publications
*For Research Use Only. Not for use in diagnostic procedures.
Product Description
Analytical Characteristics of AmplideX PCR/CE FMR1 Reagents*:
- Detects all alleles including low abundance full mutations (Figure 1)
- Accurately sizes any repeat up to 200 CGG repeats (Figure 2)
- Resolves female zygosity (Figure 3)
- Detects presence of AGG interruptions (Figure 4)
Figure 1: Amplification of Asuragen’s Methylation and Sensitivity Control which has a 5% full mutation in a background of 95% Normal
Figure 2. Female premutation sample with accurate sizing of Normal (30 CGG) and Pre mutation allele (56 CGG)
Figure 3: The difference in the “stutter” peak patterns of homozygous and heterozygous female provides a clear resolution of zygosity
Figure 4. Female Full Mutation sample with AGG interruptions as indicated by sudden decrease in peak heights of the “stutter” peak profile
Ordering
Product Name | Number of Reactions | Catalog Number |
---|---|---|
AmplideX PCR/CE FMR1 Control | 24 UL | 49513 |
AmplideX mPCR FMR1 Control | 24 UL | 49514 |
AmplideX PCR/CE FMR1 Reagents | 100 | 49402 |
AmplideX mPCR FMR1 Kit | 24 | 49442 |
AmplideX PCR/CE FMR1 Reporter | N/A | 49576 |
T 1-877-777-1874; 512-681-5200 F 512-681-5202 E orders@asuragen.com
References
- Referenced in over 30 peer reviewed publications and used in over 200 laboratories, the AmplideX® PCR/CE FMR1 Reagents* are globally recognized as best-in-class for assessment of CGG repeats in the FMR1 gene.
- Key resources
- Videos
AmplideX® PCR/CE FMR1 Kit
AmplideX PCR/CE FMR1 Kit is an in vitro diagnostic (IVD) device for use in clinical laboratories for detection of the CGG repeats in the fragile X mental retardation (FMR1) gene. The device is intended to aid in the diagnosis of fragile X syndrome and fragile X associated disorders, e.g. tremor and ataxia syndrome (FX-TAS) and primary ovarian insufficiency (FXPOI), through determination of CGG repeat length up to 200 CGG and detection of alleles greater than 200 CGG. The kit provides a PCR-only approach based on Triplet Repeat Primed PCR (TP-PCR) design to reliably amplify and detect all alleles including Full Mutations.
Features & Benefits
AmplideX PCR/CE FMR1 Kit has created an easy-to-use, accessible, high performance method for laboratories to reliably analyze CGG repeats and detect interrupting AGG sequences in the FMR1 gene.
Reduced ComplexityEase-of-analysis of the FMR1 gene has been simplified through:
- Implementation of proprietary PCR solution for amplifying GC-rich regions
- Automation of result calling using AmplideX PCR/CE FMR1 Reporter
Optimized WorkflowValuable operator hands-on time has been significantly reduced through:
- Direct injection of PCR products (no PCR clean up) in to Capillary Electrophoresis platforms
- Decreased need for Southern blot analysis (up to 50 fold)
- End-to-end solution for FMR1 analysis including all necessary reagents and software
Quality PerformancePerforming FMR1 Analysis with Greater Sensitivity and Accuracy:
- Detection of all allele expansions, including low abundance full mutation size mosaics with up to at least 1300 CGG repeats
- Up to 875 fold more sensitive than Southern blot1
- Resolution of female homozygous and heterozygous samples and indication of interrupting AGG sequences
- Proven performance as indicated by more than 30 peer reviewed publications
Product Description
Analytical Characteristics of AmplideX PCR/CE FMR1 Kit:
- Proven clinical accuracy compared to Southern Blot (Table 1)
- Detects all alleles including low abundance full mutations (Figure 1)
- Accurately sizes all alleles up to 200 CGG repeats (Figure 2)
- Resolves female zygosity (Figure 3)
- Detects presence of AGG interruptions (Figure 4)
Table 1: Diagnostic Sensitivity of 100%; Diagnostic Specificity of 98.4% and Overall Accuracy of 99%*These 2 samples presented premutation alleles by both methods and low intensity full mutation alleles detected only by the AmplideX PCR/CE FMR1 Kit
Figure 1: Amplification of Asuragen’s Methylation and Sensitivity Control which has a 5% full mutation in a background of 95% Normal
Figure 2. Female Pre-mutation sample with accurate sizing of Normal (30 CGG) and Pre-mutation allele (56 CGG)
Figure 3: The difference in the “stutter” peak patterns of homozygous and heterozygous female provides a clear resolution of zygosity
Figure 4. Female Full Mutation sample with AGG interruptions as indicated by sudden decrease in peak heights of the “stutter” peak profile
Ordering
Product Name | Number of Reactions | Catalog Number |
---|---|---|
AmplideX PCR/CE FMR1 Control* | 24 UL | 49513 |
AmplideX mPCR FMR1 Control* | 24 UL | 49514 |
AmplideX PCR/CE FMR1 Reagents* | 100 | 49402 |
AmplideX PCR/CE FMR1 Kit | 100 | 76008 |
AmplideX mPCR FMR1 Kit* | 24 | 49442 |
AmplideX PCR/CE FMR1 Reporter* | N/A | 49576 |
T 1-877-777-1874; 512-681-5200 F 1-512-681-5202 E orders@asuragen.com
References
- Referenced in over 30 peer reviewed publications and used in over 200 laboratories, the AmplideX® PCR/CE FMR1 Reagents* are globally recognized as best-in-class for assessment of CGG repeats in the FMR1 gene.
- Key resources
- Videos
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下图为ELISA原理图,夹心法即Sandwich ELISA,竞争法就是Competitive ELISA
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1. 直接法测定抗原 2.间接法测定抗体 3.双抗体夹心法测定抗原4. 竞争法测定抗原 厚百生物专业提供各类生化实验试剂、仪器、耗材,提供技术服务,满足您的实验室常规采购需求。厚百,让您更省心!
请大家帮帮忙,第一步加完抗原后必须封闭吗
目前,我用该蛋白的鼠源性单抗来包被ELISA板子,5%BSA37度封闭二小时后,洗涤四次后将板子分三个组加入抗原:其中一个组中加入的是我重组的蛋白,另一个组为用其它方法证实含该蛋白的人体内血清,最后一个组加入PBS。37度一小时后洗涤四次后每孔加入我制备的兔源性多抗。37度一小时洗涤六次后我再加入酶标记的抗兔的抗体37度40分钟后洗涤六次后TMB显色。
问题就在我显色后所有的孔都是阳性,其中以抗原为我的重组蛋白组与PBS组OD值最高,两组都可以达到1.8以上,而加入的抗原为人体血清组低,为0.6-1.0不等。
后面我优化条件,封闭液用过5%的脱脂奶粉,2%的BSA,包被的单抗浓度摸了梯度,加入的多抗也摸了梯度,酶标的抗体也摸了梯度。结果都没有改变,连趋势每次都是一样以重组蛋白组与PBS组OD最高。
今天我还试着把抗原换成其它蛋白也做出了强阳性?
请教各位高手,我的ELISA该如何往下面做了啊,是不是我的抗体本身有问题呢?
对了,我包被用的单抗是用GE公司的预装柱纯化的,很纯,在考染中是看不到一条杂带的。
(2)封闭:将已包被的板用洗涤液洗涤2次,每孔加入120 μL封闭液,封闭一段时间,取出,甩干备用。
(3)竞争反应:在制备好的板中每孔加入50 μL系列浓度的标准品溶液和50 μL稀释好的酶标抗体溶液,孵育一段时间。
(4)洗板:用洗涤液洗涤5次。
(5)显色反应:每孔加入显色溶液100 μL,孵育一段时间。
(6)终止:每孔各加入终止液,在酶标仪上测定各孔的吸光值。