
ELISA 2.0 kit for measuring Canis familiaris allergen,Can f 1. | |||||||
Antibodies: | 10D4/pAb | ||||||
Standard Range: | 50-0.1 ng/mL | ||||||
Limit of Detection: | 0.39ng/mL | ||||||
Background: | OD<0.08 at 450nm | ||||||
Coefficient of Determination: | R-squared>0.98 | ||||||
Contents: | Plate: Pre-coated with anti-Can f 1 monoclonal antibody 10D4Vial 1: (white top) Can f 1 allergen standardConcentration: 500 ng/mlVial 2: (brown) Rabbit anti Can f 1 antibodyVial 3: (blue top) Peroxidase-conjugated Goat Anti-RabbitBottle 1: Wash buffer, (10x concentrate)Bottle 2: Assay buffer, (10x concentrate)Bottle 3: TMB developing substrateBottle 4: Stop solution (0.5N sulfuric acid) | ||||||
Storage: | The ELISA 2.0 kit should be stored at 2-8°C. | ||||||
Expiry: | 6 months from date of receipt | ||||||
Not Provided: | Type I ultrapure water or 18.2MΩ de-ionized waterVolumetric measuring equipment (e.g. serological pipette, graduated cylinder)Clean containers for buffer and reagent preparationCalibrated single and multi-channel micropipettes and tipsVortex mixerPlate reader capable of reading absorbance at 450nmAnalysis software (recommended, but not required) | ||||||
Notes: | The allergen standard is recommended for immunoassay calibrationpurposes only.A list of frequently asked questions and troubleshooting guide can be found under the ‘Support’ tab on our website. | ||||||
Product Resources: | EPC-CF1-1 Certificate of Analysis Validation Performance Data | ||||||
For research and commercial use in vitro:not for human in vivo or therapeutic use. | |||||||
Certificate of Analysis | |||||||
Pre-coated Plate: | 96-well polystyrene microtiter plate coated withmonoclonal antibody 10D4 and treated with stabilizingagent. Sealed in foil pouch with desiccant. | ||||||
Monoclonal Antibody: | 10D4 (clone 10D4 C4 A10) | ||||||
Immunogen: | Can f 1 | ||||||
Isotype: | Mouse IgG2a/kappa | ||||||
Specificity: | Binds to an epitope on dog Canis familiaris allergen, Can f 1. | ||||||
Purification: | Produced in ascites and purified by affinity chromatography using Protein A. Single heavy and light chain bands on SDS-PAGE. | ||||||
Certificate of Analysis | |||||||
Detection Antibody: | Rabbit polyclonal antiserum | ||||||
Immunogen: | Dog fur extract | ||||||
Isotype | Multiple | ||||||
Specificity: | The antiserum contains IgG antibodies to dogCanis familiaris allergen, Can f 1, as well as IgG antibodies to other dog allergens.. | ||||||
Purification: | Produced in ascites and purified by affinity chromatography using Protein A. Single heavy and light chain bands on SDS-PAGE. | ||||||
Activity: | Titrated for use in ELISA at 1/1000 dilution. Prepared in 1% BSA/50% glycerol/PBS, pH 7.4, 0.22 μm filtered, preservative free. | ||||||
Certificate of Analysis | |||||||
Allergen Standard: | Purified natural Can f 1 prepared in 1% BSA/50% glycerol/PBS, pH 7.4. | ||||||
Concentration/Calibration: | 500 ng/mL (based on amino acid analysis) | ||||||
References: | |||||||
1. Larsen JN, Ford A, Gjesing B, et al. The collaborative study of the international standard of dog, Canis domesticus, hair/dander extract. J Allergy Clin Immunol 1988; 82:318-330. | |||||||
2. DeGroot H, Goei K, Van Sicter P, et al. Affinity purification of a major and minor allergen from dog extract: Serologic activity of affinity-purified Can f 1 and Can f 1-depleted extract. J Allergy Clin Immunol 1991; 87:1056-61. | |||||||
3. Schou C, Hansen G, Linter T, et al. Assay for the major dog allergen, Can f 1 investigation of house dust samples and commercial dog extracts. J Allergy Clin Immunol 1992; 88:847-53. | |||||||
4. Ingram JM, Sporik R, Rose G, Honsiger R, Chapman MD, Platts-Mills TAE. Quantitative assessment of exposure to dog (Can f 1) and cat (Fel d 1) allergens: Relationship to sensitization and asthma among children living in Los Alamos, New Mexico. J Allergy Clin Immunol 1995; 96:449-456. | |||||||
5. Sporik RB, Ingram JM, Price W, Sussman JH, Honsiger RW, Platts-Mills TAE. Association of asthma with serum IgE and skin test reactivity to allergens among children living at high altitude: Tickling the dragon’s breath. Amer J Resp Crit Care Med 1995;151:1388-1392. | |||||||
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现在国内最差也是用3代试剂,有些地方会用4代试剂。
4代试剂(检查抗原+抗体)——窗口期为4周。因为抗原于3-4周达到复制的峰值,此时通过4代试剂检查,如果感染了HIV,抗原/抗体至少有一个为阳性,如果都是阴就排除了。
3代试剂(只查抗体)——窗口期为6周。
以上为理论分析+临床经验的结果,可以说是99.9%的准确度。
但是目前FDA、CDC和试剂生产商统一达成的共识,也就是针对普通人,最保守的窗口期是3个月。无论什么试剂,3个月都100%排除。
ELISA的基础是抗原或抗体的固相化及抗原或抗体的酶标记。结合在固相载体表面的抗原或抗体仍保持其免疫学活性,酶标记的抗原或抗体既保留其免疫学活性,又保留酶的活性。在测定时,受检标本(测定其中的抗体或抗原)与固相载体表面的抗原或抗体起反应。用洗涤的方法使固相载体上形成的抗原抗体复合物与液体中的其他物质分开。再加入酶标记的抗原或抗体,也通过反应而结合在固相载体上。此时固相上的酶量与标本中受检物质的量呈一定的比例。加入酶反应的底物后,底物被酶催化成为有色产物,产物的量与标本中受检物质的量直接相关,故可根据呈色的深浅进行定性或定量分析。由于酶的催化效率很高,间接地放大了免疫反应的结果,使测定方法达到很高的敏感度。
需要的仪器有:酶标仪、恒温箱、枪、枪头、烧杯、滤纸.当然有自动洗板机最好.
复习了很多文献在测定方法里面均提到了CV
例如:
Theintra-assaycoefficientofvariationfortheassaywas4-6%.
Theanalyticcoefficientofvariationwas10.2%
问题是:
我是否可以直接使用说明书里的CV值?
如果不可以,我该如何操作进行计算
首先要什么有什么的,你得好好考虑一下。
2、看生产地址
根本没有生产地址,我们知道做实验做产品需要很多的仪器、试剂、耗材,没有人相信一间简单的屋子可以生产各种样的试剂盒。
3、看产品包装
没有任何的生产地址、联系方式等信息,这种产品有问题了连个投诉的地方都没有。
4、看公司网站
有些打着国外原装旗号,整个公司网站为英文页面,实际注册IP地址在中国。如果写着国外的地址,让你国外的朋友实地去看一下!
5、做交叉验证
拿对方提供的几个种类的试剂盒,把里面的关键组份相互替换做做实验,如果交叉严重,只能说明是一种原料生产的试剂盒贴了不同的标签。
6、看价格
价格低得离谱,却打着进口大公司原料分装,核算成本,这种低得离谱的价格是连原料都买不起的。

