CLIA Kit for Fibroblast Growth Factor 13 (FGF13)
FHF2; Fibroblast Growth Factor Homologous Factor 2
- Product No.CCC915Hu
- 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 MethodCompetitive Inhibition
- Assay Length2h
- Detection Range6.17-500pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 2.42pg/mL.
- Sample Typeserum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- DownloadInstruction Manual
- UOM48T96T96T*596T*1096T*100
- FOBUS$ 588 For more details, please contact local distributors!US$ 840 For more details, please contact local distributors!US$ 3780 For more details, please contact local distributors!US$ 7140 For more details, please contact local distributors!US$ 58800 For more details, please contact local distributors!
Specificity of the CLIA Kit for Fibroblast Growth Factor 13 (FGF13)
This assay has high sensitivity and excellent specificity for detection of Fibroblast Growth Factor 13 (FGF13).No significant cross-reactivity or interference between Fibroblast Growth Factor 13 (FGF13) and analogues was observed.
Recovery of the CLIA Kit for Fibroblast Growth Factor 13 (FGF13)
Matrices listed below were spiked with certain level of recombinant Fibroblast Growth Factor 13 (FGF13) and the recovery rates were calculated by comparing the measured value to the expected amount of Fibroblast Growth Factor 13 (FGF13) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 96-104 | 101 |
| EDTA plasma(n=5) | 97-104 | 101 |
| heparin plasma(n=5) | 97-104 | 101 |
Precision of the CLIA Kit for Fibroblast Growth Factor 13 (FGF13)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibroblast Growth Factor 13 (FGF13) 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 13 (FGF13) 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 Fibroblast Growth Factor 13 (FGF13)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Fibroblast Growth Factor 13 (FGF13) 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) | 88-95% | 79-104% | 94-103% | 85-101% |
| EDTA plasma(n=5) | 79-102% | 99-105% | 79-101% | 94-103% |
| heparin plasma(n=5) | 81-93% | 89-101% | 93-101% | 84-98% |
Stability of the CLIA Kit for Fibroblast Growth Factor 13 (FGF13)
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 Fibroblast Growth Factor 13 (FGF13)
1. Prepare all reagents, samples and standards;2. Add 50µL standard or sample to each well.And then add 50µL prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37°C;3. Aspirate and wash 3 times;4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;5. Aspirate and wash 5 times;6. Add 100µL Substrate Solution. Incubate 10 minutes at 37°C;7. Read RLU value immediately.
Test principle of the CLIA Kit for Fibroblast Growth Factor 13 (FGF13)
The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to Fibroblast Growth Factor 13 (FGF13). A competitive inhibition reaction is launched between biotin labeled Fibroblast Growth Factor 13 (FGF13) and unlabeled Fibroblast Growth Factor 13 (FGF13) (Standards or samples) with the pre-coated antibody specific to Fibroblast Growth Factor 13 (FGF13). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Fibroblast Growth Factor 13 (FGF13) in the sample. 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 reverse proportional to the Fibroblast Growth Factor 13 (FGF13) level in the sample or standard.
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Related products
| Catalog No. | Organism species: Homo sapiens (Human) | Applications (RESEARCH USE ONLY!) |
| APC915Hu02 | Active Fibroblast Growth Factor 13 (FGF13) | Cell culture; Activity Assays. |
| RPC915Hu02 | Recombinant Fibroblast Growth Factor 13 (FGF13) | Positive Control; Immunogen; SDS-PAGE; WB. |
| RPC915Hu01 | Recombinant Fibroblast Growth Factor 13 (FGF13) | Positive Control; Immunogen; SDS-PAGE; WB. |
| APC915Hu01 | Active Fibroblast Growth Factor 13 (FGF13) | Cell culture; Activity Assays. |
| CPC915Hu21 | OVA Conjugated Fibroblast Growth Factor 13 (FGF13) | Immunogen; SDS-PAGE; WB. |
| PAC915Hu08 | Polyclonal Antibody to Fibroblast Growth Factor 13 (FGF13) | WB; IHC; ICC; IP. |
| PAC915Hu01 | Polyclonal Antibody to Fibroblast Growth Factor 13 (FGF13) | WB; IHC; ICC; IP. |
| PAC915Hu02 | Polyclonal Antibody to Fibroblast Growth Factor 13 (FGF13) | WB; IHC; ICC; IP. |
| LAC915Hu71 | Biotin-Linked Polyclonal Antibody to Fibroblast Growth Factor 13 (FGF13) | WB; IHC; ICC. |
| LAC915Hu81 | FITC-Linked Polyclonal Antibody to Fibroblast Growth Factor 13 (FGF13) | WB; IHC; ICC; IF. |
| MAC915Hu21 | Monoclonal Antibody to Fibroblast Growth Factor 13 (FGF13) | WB; IHC; ICC; IP. |
| MAC915Hu22 | Monoclonal Antibody to Fibroblast Growth Factor 13 (FGF13) | WB; IHC; ICC; IP. |
| CEC915Hu | ELISA Kit for Fibroblast Growth Factor 13 (FGF13) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| HEC915Hu | High Sensitive ELISA Kit for Fibroblast Growth Factor 13 (FGF13) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| SCC915Hu | CLIA Kit for Fibroblast Growth Factor 13 (FGF13) | Chemiluminescent immunoassay for Antigen Detection. |
| CCC915Hu | CLIA Kit for Fibroblast Growth Factor 13 (FGF13) | Chemiluminescent immunoassay for Antigen Detection. |
| KSC915Hu11 | ELISA Kit DIY Materials for Fibroblast Growth Factor 13 (FGF13) | Main materials for "Do It (ELISA Kit) Yourself". |
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一.样本种属来源:
首先要考虑的就是实验样本来源于什么物种。
1、哺乳动物的组织或者细胞样本,通常选择β-actin、β-tubulin、GAPDH、Lamin B、Histone H3、Na,K atpase等。
2、植物来源实验样本,则可以选择plantactin、Rubisco等。
3、其他来源样本研究较少,所以就应该参照文献报导,选择合适的蛋白作为内参。
二.目的蛋白分子量:
选择内参抗体时,应该考虑目的蛋白分子量的大小。通常应该保证目的蛋白与内参蛋白分子量相差5KD以上。比如目的蛋白分子量为45KD,此时不适宜选择β-actin作为内参,可以考虑选择GAPDH或者β-tubulin作为内参。
三.目的蛋白表达部位:
就一般的蛋白检测来说,β-actin、β-Tubulin抗体等就可以了,而针对于核蛋白的定量,特别是样本蛋白就是核蛋白时,选择恰当的核蛋白内参则更能体现内部参照的价值。常用的核内参抗体有Lamin A、Lamin B、Histone H3,除此之外,其它常见的核蛋白内参还有PCNA、K70、K80等,在一些文献报道中,Erk2、TATA binding protein(TBP)以及c-Jun、c-Fos等都有使用。而对于膜蛋白检测,常用的内参抗体为Na,K ATPase。对于线粒体蛋白的检测,常用VDAC1和COX IV作为内参抗体。
以上几条原则只是针对通常情况,但是需要注意的问题是——内参的选择需要考虑实际的试验环境,比如某些细胞中,由于组织缺氧、糖尿病等因素会导致GAPDH的表达增高,不适合做内参。比如在涉及细胞增殖相关试验中,c-Jun由于自身表达变化就不适合做内参;而在凋亡实验时,TBP、Lamin等也不适合作为内参。因此设计实验方案的时候应该考虑这些因素并查询相应文献,在实验过程中也应该注意如果内参表达出现异常,应考虑这方面因素。
β-Actin作为内参是得到了公认的,这是针对大多数组织和细胞来说的,它广泛分布于细胞浆内,表达量非常丰富。Beta-actin由375个氨基酸组成,分子量大小为42-43kDa左右。
β-actin的蛋白水平通常不会发生改变,因此被广泛用于Western时上样量是否一致的参照,也常被用于免疫染色观察细胞的微丝结构。在用作Western的参照时,Actin抗体和Tubulin抗体的主要不同之处在于两者所识别蛋白的分子量不同,这样可以选择合适的参照在同一块胶同一张膜上实现同时检测目标蛋白和参照蛋白。向左转|向右转
内参抗体:
Beta-Actin mAb (1C7) 细胞总蛋白 42 kD Abbkine A01010 小鼠源单克隆
Beta-Actin mAb (1C7) , HRP 细胞总蛋白 42 kD Abbkine A01015 小鼠源单克隆,HRP偶联
GAPDH mAb (2B5) 细胞总蛋白 36 kD Abbkine A01020 小鼠源单克隆
GAPDH mAb (2B5) , HRP 细胞总蛋白 36 kD Abbkine A01025 小鼠源单克隆,HRP偶联
Beta-Tubuline mAb (3G6) 细胞总蛋白 55 kD Abbkine A01030 小鼠源单克隆
Plant Actin mAb (3T3) 植物细胞总蛋白 42 kD Abbkine A01050 小鼠源单克隆
PCNA mAb (1D7) 细胞核蛋白 28 kD Abbkine A01040 小鼠源单克隆
COX IV mAb (14Y2) 细胞线粒体总蛋白 16 kD Abbkine A01060 小鼠源单克隆
Histone H3 mAb (2D10) 细胞核蛋白 18 kD Abbkine A01070 小鼠源单克隆
标签抗体(Tag Antibody)可用于检测各种商品化表达载体上的标签序列(如:Myc、Flag、His、GST、HA等),籍以分析目的蛋白的表达含量及其功能;
标签抗体:
Anti-Biotin Antibodies
Anti-Dye Antibodies
Anti-FITC Antibodies
Anti-Fluorescent Protein Antibodies
Anti-HRP Antibodies
Beta Galatosidase Antibodies
FLAG Tag Antibodies
GST Tag Antibodies
HA Tag Antibodies
His Tag Antibodies
Myc Tag Antibodies
TAP Tag Antibodies
V5 Tag Antibodies
一般可与大鼠、小鼠、人、兔均有交叉反应,即可作为上述任一来源蛋白的内参。
Actin分子量为42kD,为肌动蛋白
GAPDH分子量为36kD,为磷酸脱氢酶
因为内参有好几种,分子量差别也比较大,更换分子量与检测蛋白差距更大的内参就可以避免这个问题
另外也可以先用一抗孵育显色和检测,再用Strip缓冲液洗掉膜上的抗体,重新进行内参的抗体孵育显色检测。这样也可以将检测蛋白和内参显色在同一张膜上
所以免疫组化在组织定位和定性上比较准确,但是在定量上不够准确。
western blot也是利用抗体和抗原之间的结合具有高度的特异性。经过聚丙烯酰胺凝胶电泳分离的蛋白质样品,转移到固相载体上,固相载体以非共价键形式吸附蛋白质,且能保持电泳分离的多肽类型及其生物学活性不变。以固相载体上的蛋白质或多肽作为抗原,与对应的抗体起免疫反应,再与酶或同位素标记的第二抗体起反应。
western blot因为有内参标定,所以在定量上要更加准确。但是在组织定位上不如免疫组化
GAPDH
万能内参
主要看你的目标蛋白的分子量多大,“目标蛋白要与内参蛋白的分子量差异大!”
方便检测
核蛋白,有很多种,分子量不同
膜蛋白,有很多很多种,分子量不同
浆蛋白,有成千上万种,分子量不同。
具体实验,具体目的蛋白,具体的内参。
虽然没有具体用过检测这个抗原的抗体,但是如果希望抗体特异性好点,能检测丰度低的蛋白,还是选Abcam吧,Santa有时候全凭人品了,内参抗体什么的还能凑活,其他的比较勉强。


