Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine®

alpha 2-Macroglobulin ELISA kit for Human

Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine™ (96 Tests). Quantitate Human A2M in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 20 pg/ml. Cited in 2 publication(s). Independently reviewed in 1 review(s).

Product Info Summary

SKU: EK1118
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Product Name

Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine®

View all alpha 2-Macroglobulin ELISA kits

SKU/Catalog Number

EK1118

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine™ (96 Tests). Quantitate Human A2M in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 20 pg/ml.

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cite This Product

Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1118)

Clonality of Antibodies

See Datasheet for details

Immunogen

Target protein isolated from human plasma

Sensitivity

<20 pg/ml

Assay Range

625 pg/ml - 40,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK1118 for more details

Cross-reactivity

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

Reactive Species

EK1118 is reactive to A2M in Human samples

Validated Sample Types

cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Application Guarantee

EK1118 is guaranteed for ELISA in Human by Boster Guarantee

See how Boster Bio validate our ELISA kits: ELISA Validation Information

Background of alpha 2-Macroglobulin

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.

Kit Components

Catalog Number Description Quantity
EK1118-CAP Anti-Human A2M Pre-coated 96-well strip microplate 1
EK1118-ST Human A2M Standard 2 vials, 40 ng/tube
EK1118-DA Human A2M Biotinylated antibody (100x) 100ul
AR1103 Avidin-Biotin-Peroxidase Complex (100x) 100ul
AR1106-1 Sample Diluent 30ml
AR1106-2 Antibody Diluent 12ml
AR1106-3 Avidin-Biotin-Peroxidase Diluent 12ml
AR1104 Color Developing Reagent (TMB) 10ml
AR1105 Stop Solution 10ml
AR1106-5 Wash Buffer (25x) 20ml
PLA-SEA Adhesive plate sealers 4

*The kit components are not available for individual purchase.

Materials Required But Not Included In Kit

  • Microplate Reader capable of reading absorbance at 450nm.
  • Incubator.
  • Automated plate washer (optional).
  • Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
  • Multichannel pipettes are recommended for large amount of samples.
  • Deionized or distilled water.
  • 500ml graduated cylinders.
  • Test tubes for dilution.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)0625125025005000100002000040000
O.D.0.0220.0890.1470.2540.4520.7891.3061.892

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect alpha 2-Macroglobulin, Dilution ratio of 1:200000, concentration in serum and plasma is 0.63-1.7mg/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)13673876267841316394226889
Standard deviation97.05236.432062.36103.96240.462151.12
CV (%)7.1%6.1%7.7%7.9%6.1%8%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of alpha 2-Macroglobulin in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 11367128514691391137865.614.7%
Sample 238763671326033213532253.067.1%
Sample 326784320342963029138293961864.336.3%
*number of samples for each test n=16.

Gene/Protein Information For A2M (Source: Uniprot.Org, NCBI)

Gene Name

A2M

Full Name

Alpha-2-macroglobulin

Weight

163.291kDa

Superfamily

protease inhibitor I39 (alpha-2-macroglobulin) family

Alternative Names

A2M; alpha 2Macroglobulin; alpha 2-Macroglobulin; alpha-2-M; alpha-2-macroglobulin; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5; CPAMD5; CPAMD5DKFZp779B086; FWP007; S863-7 A2M A2MD, CPAMD5, FWP007, S863-7 alpha-2-macroglobulin alpha-2-macroglobulin|C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5|alpha-2-M

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

For more info on A2M, check out the A2M Infographic

A2M infographic

We have 30,000+ of these available, one for each gene! check them out.

In this infographic you will see the following information for A2M: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].

Hello CJ!

EK1118 has been cited in 2 publications:

*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.

Disequilibrium of Blood Coagulation and Fibrinolytic System in Patients With Coronary Artery Ectasia

Neutrophil serine proteases and their endogenous inhibitors in coronary artery ectasia patients

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1 Reviews For Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine®

0

Reasonable sensitivity and reproducibility--Preethi Vijayaraj, UCLA, Pediatrics, Postdoctoral Fellow

Excellent

Source: Biocompare.com

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.")

Verified customer

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8 Customer Q&As for Human A2M/Alpha 2-Macroglobulin ELISA Kit PicoKine®

Question

Q: is it okay to use citrate plasma as samples in Human alpha 2-Macroglobulin Picokine® ELISA Kit (Catalog # EK1118)?

R. Lee

Verified customer

Asked: 2020-11-09

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of alpha 2-Macroglobulin causing disruption of its protein structure. alpha 2-Macroglobulin may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes needed for detection and block the antigen antibody reaction. We have tested the alpha 2-Macroglobulin ELISA, treating samples with various anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2020-11-09

Question

I will be using samples that are pre-anticoagulated and the anticoagulation is performed with ATC. However, the protocol states that sodium citrate, EDTA or heparin can be used with these Human A2M/Alpha2-Macroglobulin ELISA Kit. So, should I perform another coagulation step with one of the above?

Verified Customer

Verified customer

Asked: 2019-12-31

Answer

The antibody Human A2M/Alpha2-Macroglobulin ELISA Kit have not been tested on ATC samples, thus we cannot give you further recommendation about it.

Boster Scientific Support

Answered: 2019-12-31

Question

Q: how can I thaw whole blood sample for alpha 2-Macroglobulin ELISA after freezing?

Verified Customer

Verified customer

Asked: 2019-06-09

Answer

A: we do not recommend freezing and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test alpha 2-Macroglobulin for a later time, you should let the blood clot in glass tubes and separate the serum to freeze for later analysis.

Boster Scientific Support

Answered: 2019-06-09

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for analysis alpha 2-Macroglobulin using the EK1118 Human alpha 2-Macroglobulin Picokine® ELISA Kit?

B. Babu

Verified customer

Asked: 2018-10-20

Answer

A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may change the stabilityof samples. It is recommend to assay sample on priority basis collection when possible, or aliquot into single use volumes and store samples frozen. avoid repetitive freeze-thaw cycles with the stored samples to prevent protein degradation.

Boster Scientific Support

Answered: 2018-10-20

Question

Q: if the enzyme conjugated alpha 2-Macroglobulin antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the alpha 2-Macroglobulin antigen?

Verified Customer

Verified customer

Asked: 2018-05-23

Answer

A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.

Boster Scientific Support

Answered: 2018-05-23

Question

Q: can you suggest the dilution ratio of serum samples for detection of alpha 2-Macroglobulin in Human serum? I am trying to measure a few parameters and it requires 100ul of diluted samples for each well. We have low serum volumes so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2018-01-04

Answer

A: unable to know the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the alpha 2-Macroglobulin ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2018-01-04

Question

What volume should the standard be reconstituted with Human A2M/Alpha2-Macroglobulin ELISA Kit?

Verified Customer

Verified customer

Asked: 2015-11-24

Answer

For Human A2M/Alpha2-Macroglobulin ELISA Kit, the standard vial contains 160ug of human alpha-2-macroglobulin, so to make a stock solution 40ug/mL, you need to reconstitute the standard in 4 mL of 1x diluent.

Boster Scientific Support

Answered: 2015-11-24

Question

For EK1118, could you confirm the immunogen/expression system information? On the datasheet, it says "Expression system for standard: from human plasma"

Verified Customer

Verified customer

Asked: 2014-02-06

Answer

Yes, the information on the datashet is correct. Expression system for standard: from human plasma

Boster Scientific Support

Answered: 2014-02-06

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