Anti-alpha 2 Macroglobulin antibody (ab84176)

Overview

  • Product nameAnti-alpha 2 Macroglobulin antibodySee all alpha 2 Macroglobulin primary antibodies ...
  • Description
    Rabbit polyclonal to alpha 2 Macroglobulin
  • Tested applicationsWB, ICC/IF more details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1450 to the C-terminus of Human alpha 2 Macroglobulin.

    (Peptide available as ab93750.)

  • Positive control
    • This antibody gave a positive signal in both human plasma total protein and human placenta tissue lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab84176 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 185 kDa (predicted molecular weight: 163 kDa).Can be blocked with Human alpha 2 Macroglobulin peptide (ab93750).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • FunctionIs 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.
  • Tissue specificitySecreted in plasma.
  • Sequence similaritiesBelongs to the protease inhibitor I39 (alpha-2-macroglobulin) family.
  • Developmental stageContrary to the rat protein, which is an acute phase protein, this protein is always present at high levels in circulation.
  • Cellular localizationSecreted.
  • Target information above from: UniProt accession P01023 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • A2M antibody
    • A2MG_HUMAN antibody
    • Alpha 2 M antibody
    • Alpha 2M antibody
    • Alpha-2-M antibody
    • Alpha-2-macroglobulin antibody
    • C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5 antibody
    • CPAMD5 antibody
    • DKFZp779B086 antibody
    • FWP007 antibody
    • S863 7 antibody
    see all

Anti-alpha 2 Macroglobulin antibody images

  • All lanes : Anti-alpha 2 Macroglobulin antibody (ab84176) at 1 µg/ml

    Lane 1 : Human Plasma Total Protein Lysate
    Lane 2 : Placenta (Human) Tissue Lysate - adult normal tissue (ab29745)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 163 kDa
    Observed band size : 185 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 48 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 15 minutes
  • ICC/IF image of ab84176 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab84176, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 1µg/ml, and in 100% methanol fixed (5 min) MCF7 cells at 5µg/ml.

References for Anti-alpha 2 Macroglobulin antibody (ab84176)

ab84176 has not yet been referenced specifically in any publications.

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http://www.abcam.com/alpha-2-macroglobulin-antibody-ab84176.html

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Read More
Application Western blot
Sample Human Tissue lysate - whole (human heart left ventricular)
Loading amount 40 µg
Specification human heart left ventricular
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dawn Bowles

Verified customer

Submitted Jan 10 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"