|Size||96wells/kit, with removable strips.|
|Sample Type||cell culture supernates, cell lysates, serum and plasma (heparin, EDTA).|
|Product Name||Human A2M/Alpha2-Macroglobulin PicoKine™ ELISA Kit|
|Storage & Handling||Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)|
|Size||96wells/kit, with removable strips.|
|Description||Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human A2M/alpha2-Macroglobulin. 96wells/kit, with removable strips.|
|Cite This Product||Human A2M/Alpha2-Macroglobulin PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1118)|
|Sample Type||cell culture supernates, cell lysates, serum and plasma (heparin, EDTA)..
Anticoagulant(s): heparin or EDTA
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants that are not specified above or contact us to check for feasibility.
|Immunogen||Expression system for standard: from human plasma|
|Cross Reactivity||There is no detectable cross-reactivity with other relevant proteins.|
|Antibody Clonalities||Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|polyclonal antibody from goat
|EK1118-CAP||96-well plate precoated with anti-Human A2M antibody||1|
|EK1118-ST||lyophilized recombinant Human A2M standard||40ng/tube|
|EK1118-DA||biotinylated anti-Human A2M antibody||130ul|
|AR1106-1||sample diluent buffer||30ml|
|AR1106-2||antibody diluent buffer||12ml|
|AR1106-3||ABC diluent buffer||12ml|
|AR1104||TMB color developing agent||10ml|
|AR1105||TMB stop solution||10ml|
*Why there is no wash buffer? Our Avidin-Biotin-Peroxidase Diluent contains the detergent (TWEEN) normally present in other companies' ELISA kits. This saves you the step of having to wash with the special wash buffer and achieve similar or better signal to noise ratio. The wash can use regular wash buffers (PBS, TBS etc.) commonly found in labs.
Materials Required But Not Provided
- Microplate reader in standard size.
- Automated plate washer.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- Washing buffer (neutral PBS or TBS).
- Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
Typical Data Obtained from Human A2M/Alpha2-Macroglobulin PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-25min)
Intra/Inter Assay Precision
|Intra-Assay Precision||Inter-Assay Precision|
Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.
To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.
|Lots||Lot1 (pg/ml)||Lot2 (pg/ml)||Lot3 (pg/ml)||Lot4 (pg/ml)||Mean (pg/ml)||Standard Deviation||CV (%)|
*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.
Protein Target Info (Source: Uniprot.org)
|Tissue Specificity||Secreted in plasma. .|
|Alternative Names||Alpha-2-macroglobulin;Alpha-2-M;C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5;A2M;CPAMD5;FWP007;|
|Subcellular Localization||Secreted .|
|Molecular Weight||163291 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Is able to inhibit all four classes of proteinases by a unique 'trapping' mechanism. This protein has a peptide stretch, called the 'bait region' which contains specific cleavage sites for different proteinases. When a proteinase cleaves the bait region, a conformational change is induced in the protein which traps the proteinase. The entrapped enzyme remains active against low molecular weight substrates (activity against high molecular weight substrates is greatly reduced). Following cleavage in the bait region a thioester bond is hydrolyzed and mediates the covalent binding of the protein to the proteinase.|
|Research Areas||Cell Biology, Complement, Immunology, Innate Immunity, Protease Inhibitors, Proteolysis / Ubiquitin
*You can search these to find other products in these research areas.
|Background||Alpha-2-macroglobulin, also known as A2M or CPAMD5 is a large plasma protein found in the blood. This gene is mapped to 12p13.31. Alpha-2-macroglobulin is a protease inhibitor and cytokine transporter. It inhibits many proteases, including trypsin, thrombin and collagenase. A2M is implicated in Alzheimer disease(AD) due to its ability to mediate the clearance and degradation of A-beta, the major component of beta-amyloid deposits. This gene is able to inhibit all four classes of proteinases by a unique 'trapping' mechanism. This protein has a peptide stretch, called the 'bait region' which contains specific cleavage sites for different proteinases. When a proteinase cleaves the bait region, a conformational change is induced in the protein which traps the proteinase. The entrapped enzyme remains active against low molecular weight substrates(activity against high molecular weight substrates is greatly reduced). Following cleavage in the bait region a thioester bond is hydrolyzed and mediates the covalent binding of the protein to the proteinase.|
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Guaranteed product quality
We promise all of our products perform as described in datasheets.
- Review by Preethi Vijayaraj
"Reasonable sensitivity and reproducibility"
--Preethi Vijayaraj, UCLA, Pediatrics, Postdoctoral Fellow
Application ELISA Sample Cell culture supernatant Detection --
"Our research focuses on in vitro modeling of organ fibrosis and drug discovery. The protocol for this product was easy to perform. However, there was some amount of variability with the replicates. Was effective in detecting A2M levels at pg levels. Reasonable sensitivity and reproducibility. Easy to use." (Customer’s Tip: "Follow protocol to the T.")
1. Diluent the samples with the provided sample diluent buffer into 100ul.
2. Add 50ul of standard solution, when 50ul of sample will be added into a well.
• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
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