Human A2M/alpha2-Macroglobulin PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human A2M in cell culture supernates, cell lysates, tissue homogenates, serum and plasma (heparin, EDTA).

spezie

Human

Cross Reaction

There is no detectable cross-reactivity with other relevant proteins.

detection target

human A2M
Protein name: Alpha-2-macroglobulin
Gene Full Name: alpha-2-macroglobulin
abbreviation: A2M
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.
ID:P01023

alias

C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5|A2m|A2MD|A2MG_HUMAN|Alpha-2-M|Alpha-2-macroglobulin|C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5|CPAMD5|FWP007|Pregnancy zone protein|S863-7

Origin

Nativ human A2M

Antibodies

a monoclonal antibody from mouse|a biotinylated polyclonal antibody from goat

Immunogene

from human plasma

sensitivity

<20pg/ml

detection range

625pg/ml-40000pg/ml

pack size

1x96 Wells

Reproducibility

|Intra-Assay Precision|Inter-Assay Precision
Sample|1|2|3|1|2|3
n|16|16|16|24|24|24
Mean(ng/ml)|6.18|12.04|25.21|6.72|13.68|26.81
Standard deviation|0.377|0.771|0.174|0.457|0.985|2.12
CV(%)|6.1|6.4|6.9|6.8|7.2|7.9

TMB(tetramethylbenzidine) incubation time

15-20

storage and Stability

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

Principle of the Elisa

Boster′s 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 Step 1

Aliquot 0.1ml per well of the 40000pg/ml, 20000pg/ml, 10000pg/ml, 5000pg/ml, 2500pg/ml, 1250pg/ml, 625pg/ml human A2M standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See ﾓSample Dilution Guidelineﾔ above for details. It is recommended that each human A2M standard solution and each sample be measured in duplicate.

tipical result

Concentration(pg/ml)|0|625|1250|2500|5000|10000|20000|40000 O.D.|0.075|0.129|0.203|0.332|0.575|1.108|1.477|2.135

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human A2M antibody|1
ST0000-40|Lyophilized recombinant human A2M standard|40ng/tube�2
AR1107|Biotinylated anti- human A2M antibody|130µl(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex (ABC)|130µl(dilution 1:100)
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
-|Adhesive cover|4