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Scientific Background
This ELISA kit applies to the in vitro quantitative determination of Human Aβ1-42 concentrations in serum, plasma and other biological fluids.
Assay Principle
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human Aβ1-42. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human Aβ1-42 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human Aβ1-42, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human Aβ1-42. You can calculate the concentration of Human Aβ1-42 in the samples by comparing the OD of the samples to the standard curve.
Performance Specifications
| Sensitivity | 9.38 pg/mL |
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| Detection Range | 15.63-1000 pg/mL |
| Total Assay Time | 3 h 30 min |
| Compatible Sample Types | Serum, plasma and other biological fluids |
| Species Reactivity | Human |
| Detection Method | Sandwich |
| Precision (CV) | Both intra-CV and inter-CV are < 10%. |
| Recovery Rate | 80%-120% |
| Storage | 2-8℃,12 months |
✓ Research-Grade Validation
Specificity
This kit recognizes Human Aβ1-42 in samples. No significant cross-reactivity or interference between Human Aβ1-42 and analogues was observed
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with Serum, plasma and other biological fluids. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 9.38 pg/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3 h 30 min, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant Aβ1-42 standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
In mammals, the APP gene is located on chromosome 21 and has a total of 8 subtypes, whose subtypes generally contain 365~770 amino acid residues. The most common are APP695, APP751, and APP770, of which APP695 is highly expressed in the central nervous system. APP is a type I transmembrane glycoprotein with a molecular weight of about 110~130 kDa, with a large extracellular (amino terminal) domain and a small cytoplasmic tail region (intracellular carboxyl terminal). APP is metabolized mainly through two pathways: amyloid (β) pathway and non-amyloid (α) pathway. The amyloid pathway is that APP is cleaved by β-secretase into sAPPβ and a C-terminal fragment containing 99 amino acids. The latter is further cleaved by γ-secreting enzymes into Aβ and ACID, and the Aβ generated by this pathway accounts for 90% to 95% of the total Aβ, including two types of Aβ1-40 and Aβ1-42, of which the content of Aβ1-40 is higher, but Aβ1-42 has A strong hydrophobic effect, its toxicity is greater, and it is easy to polymerization. These Aβ fragments accumulate in mitochondria, lysosomes, and the endoplasmic reticulum, causing these suborganelles to malfunction. In the non-starchy source pathway based on alpha secretase, the alpha secretase cuts APP into sAPP and a C-terminal fragment containing 83 amino acids, which is then cut by gamma secretase to produce P3 and ACID, but these small fragments are cleared by neurons. The mutation of APP gene can cause abnormal protein expression or hydrolysis change, thus affecting the content and composition of Aβ in cells. Aβ mainly exists in the brain in the form of Aβ40 and Aβ42, and the content of Aβ42 is low (< 10%), but easy to aggregate, and then fibrosis and deposition, thus forming a diffuse senile plaque, which is also one of the main pathological characteristics of AD. Moreover, Aβ produced by APP after cleavage by β- and γ-secretory enzymes on the cell membrane can cause oxidative stress, calcium ion inflow, and then damage mitochondria, leading to nerve cell disorders, activation of apoptosis-related proteins and factors, and finally start the apoptosis process of cells. In addition, Aβ can also indirectly cause neuronal apoptosis by causing inflammation in the brain and neurofibrillary tangles, which is an important reason for the formation and development of AD.
Tissue samples may contain endogenous or exogenous proteases during processing, leading to degradation of extracted proteins. Therefore, it's necessary to add protease inhibitors during processing to ensure the integrity of target proteins. If customers can keep samples cold and handle them quickly during processing, omitting the protease inhibitor may not have a significant effect. After preparation, samples should be tested promptly or immediately aliquoted and frozen at -20°C or -80°C.
What is the range of enzyme activity of your IL-2 freeze-dried powder
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