ELISA 2.0 kit for measuring Anacardium occidentale allergen, Ana o 3. | |||||||
| Antibodies: | 1H4/4A11 | ||||||
| Standard Range: | 40-0.08 ng/mL | ||||||
| Limit of Detection: | 0.16 ng/mL | ||||||
| Background: | OD<0.08 at 450nm | ||||||
| Coefficient of Determination: | R-squared>0.98 | ||||||
| Contents: | Plates: Pre-coated with anti-Ana o 3 monoclonal antibody 1H4Vial 1: (white top) Ana o 3 allergen standardConcentration: 400 ng/mlVial 2: (brown) anti-Ana o 3 antibody 4A11Vial 3: (blue top) Streptavidin-PeroxidaseBottle 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-AO3-5 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 plates coated withmonoclonal antibody 1H4 and treated with stabilizingagent. Sealed in foil pouch with desiccant. | ||||||
| Monoclonal Antibody: | 1H4 | ||||||
| Immunogen: | Ana o 3 | ||||||
| Isotype: | Mouse IgG2a/kappa | ||||||
| Specificity: | Binds to species specific epitope present on Anacardium occidentale allergen, Ana o 3. | ||||||
| Purification: | Produced in vitro and purified by affinity chromatography using Protein A. Single heavy and light chain bands on SDS-PAGE. | ||||||
| Certificate of Analysis | |||||||
| Detection Antibody: | 4A11 | ||||||
| Immunogen: | Ana o 3 | ||||||
| Isotype | Mouse IgG2b/kappa | ||||||
| Specificity: | Binds to species specific epitope present onAnacardium occidentaleallergen, Ana o 3. | ||||||
| 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 recombinant Ana o 3 prepared in 1% BSA/50%glycerol/PBS, pH 7.4. | ||||||
| Concentration/Calibration: | 400 ng/mL (based on amino acid analysis) | ||||||
| References: | |||||||
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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.
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。

