alpha 2 Macroglobulin ELISA Kit

Catalog No. ABIN1672904

This Colorimetric ELISA kit is designed for the quantitative measurement of Human alpha 2 Macroglobulin. There are 2 publications available.

Quick Overview for alpha 2 Macroglobulin ELISA Kit (ABIN1672904)

Target: alpha 2 Macroglobulin (A2M) (alpha-2-Macroglobulin (A2M))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 625-40.000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Product Details alpha 2 Macroglobulin ELISA Kit

Minimum Detection Limit: 625 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human A2M/alpha2-Macroglobulin

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: from human plasma

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <20pg/mL

Material not included: 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

Immunogen: from human plasma

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Belongs to the protease inhibitor I39 (alpha-2- macroglobulin) family.

Tissue Specificity: Secreted in plasma. .

Plate: Pre-coated

Protocol: human A2M ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for A2M has been precoated onto 96-well plates. Standards(from human plasma) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for A2M is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human A2M amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 40,000pg/mL, 20,000pg/mL, 10,000pg/mL, 5,000pg/mL, 2,500pg/mL, 1,250pg/mL, 625pg/mL human A2M standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human A2M standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(ng/ml): 6.18, Standard deviation: 0.377, CV(%): 6.1
• Sample 2: n=16, Mean(ng/ml): 12.04, Standard deviation: 0.771, CV(%): 6.4
• Sample 3: n=16, Mean(ng/ml): 25.21, Standard deviation: 0.174, CV(%): 6.9,
• Sample 1: n=24, Mean(ng/ml): 6.72, Standard deviation: 0.457, CV(%): 6.8
• Sample 2: n=24, Mean(ng/ml): 13.68, Standard deviation: 0.985, CV(%): 7.2
• Sample 3: n=24, Mean(ng/ml): 26.81, Standard deviation: 2.12, CV(%): 7.9

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References for alpha 2 Macroglobulin ELISA Kit (ABIN1672904)

Liu, Chen, Wu, Chen, Zhang: "Neutrophil serine proteases and their endogenous inhibitors in coronary artery ectasia patients." in: Anatolian journal of cardiology, Vol. 16, Issue 1, pp. 23-8, (2016) (PubMed).

Wu, Liu, Chen, Chen, Zhang: "Disequilibrium of Blood Coagulation and Fibrinolytic System in Patients With Coronary Artery Ectasia." in: Medicine, Vol. 95, Issue 8, pp. e2779, (2016) (PubMed).

Target Details for alpha 2 Macroglobulin

Target: alpha 2 Macroglobulin (A2M) (alpha-2-Macroglobulin (A2M))

Alternative Name: A2M

Background:

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.

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.

Synonyms: Alpha-2-macroglobulin,Alpha-2-M,C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5,A2M,CPAMD5,FWP007,

Full Gene Name: Alpha-2-macroglobulin

Cellular Localisation: Secreted.

Gene ID: 2

UniProt: P01023

Pathways: Lipid Metabolism