Overview

  • Product name
    Anti-alpha 2 Macroglobulin antibody
    See all alpha 2 Macroglobulin primary antibodies
  • Description
    Mouse polyclonal to alpha 2 Macroglobulin
  • Host species
    Mouse
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Mouse
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide: KAITYLNTGY QRQLNYKHRD GSYSTFGDKP GRSHANTWLT , corresponding to amino acids 1003/1042 of Mouse alpha 2 Macroglobulin

  • General notes


    This antibody was raised by a genetic immunization technique. Genetic immunization can be used to generate antibodies by directly delivering antigen-coding DNA into the animal, rather than injecting a protein or peptide (Tang et al. PubMed: 1545867; Chambers and Johnston PubMed 12910245; Barry and Johnston PubMed: 9234514). The animal's cells produce the protein, which stimulates the animal's immune system to produce antibodies against that particular protein. A vector coding for a partial fusion protein was used for genetic immunisation of a mouse and the resulting serum was tested in Western blot against an E.coli lysate containing that partial fusion protein. Genetic immunization offers enormous advantages over the traditional protein-based immunization method. DNA is faster, cheaper and easier to produce and can be produced by standard techniques readily amenable to automation. Furthermore, the antibodies generated by genetic immunization are usually of superior quality with regard to specificity, affinity and recognizing the native protein

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Constituents: 50% Glycerol, Whole serum
  • Purity
    Whole antiserum
  • Primary antibody notes
    This antibody was raised by a genetic immunization technique. Genetic immunization can be used to generate antibodies by directly delivering antigen-coding DNA into the animal, rather than injecting a protein or peptide (Tang et al. PubMed: 1545867; Chambers and Johnston PubMed 12910245; Barry and Johnston PubMed: 9234514). The animal's cells produce the protein, which stimulates the animal's immune system to produce antibodies against that particular protein. A vector coding for a partial fusion protein was used for genetic immunisation of a mouse and the resulting serum was tested in Western blot against an E.coli lysate containing that partial fusion protein. Genetic immunization offers enormous advantages over the traditional protein-based immunization method. DNA is faster, cheaper and easier to produce and can be produced by standard techniques readily amenable to automation. Furthermore, the antibodies generated by genetic immunization are usually of superior quality with regard to specificity, affinity and recognizing the native protein
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab52651 in the following tested applications.

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

Application Abreviews Notes
WB Use at an assay dependent dilution. Predicted molecular weight: 163 kDa. This antibody has been tested in Western blot against an E.coli lysate containing the partial recombinant fusion protein used as an immunogen. We have no data on detection of endogenous protein.

Target

  • 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.
  • Tissue specificity
    Secreted in plasma.
  • Sequence similarities
    Belongs to the protease inhibitor I39 (alpha-2-macroglobulin) family.
  • Developmental stage
    Contrary to the rat protein, which is an acute phase protein, this protein is always present at high levels in circulation.
  • Cellular localization
    Secreted.
  • 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

Images

  • All lanes : Anti-alpha 2 Macroglobulin antibody (ab52651) at 1/1000 dilution

    Lane 1 : (Left) a total protein extract from E coli with 50ng to 100 ng of a tag fusion protein of an irrelevant antigen
    Lane 2 : (Right) a total protein extract from E coli with 50ng to 500ng of the antigen (tag-antigen fusion protein)

    Secondary
    All lanes : Rabbit anti-mouse IgG + IgM, (H+L) horseradish peroxidase conjugated

    Predicted band size: 163 kDa



    The molecular weight of the band on the western blot does not correspond to the molecular weight of the natural protein because only a fragment of the gene is used and it is tagged.

References

This product has been referenced in:
  • de Castro Brás LE  et al. Plasma fractionation enriches post-myocardial infarction samples prior to proteomics analysis. Int J Proteomics 2012:397103 (2012). Read more (PubMed: 22778955) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain)
Specification
Brain

Abcam user community

Verified customer

Submitted Apr 18 2016

Question
Answer

Thank you for taking time to complete our questionnaire. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab52651:

1. I would consider altering the blocking conditions used. When testing our antibodies, our lab uses 5% BSA as a blocking reagent, so I recommend switching to this instead of milk. For unknown reasons some antibodies give stronger, more specific signals on blots blocked with BSA instead of milk, so doing this may improve the results you are seeing, and reduce the non-specific bands. An example of the above is the western blot obtained with the Abcam GAPDH antibody ab9385 : https://www.abcam.com/gapdh-antibody-hrp-loading-control-ab9385.html#GAPDH-Primary-antibodies-ab9385-3.jpg (or use the following: https://www.abcam.com/ab9385).BSA should then also be used in the dilutionbuffer.

2. I would introduce 0.1% Tween 20 into your diluent buffers and was buffers as this can help reduce the non-specific staining observed.

3. I would also suggest performing a "no primary" control to ascertain how much of the non-specificty observed is due to the secondary antibody. This is particularly important as the antibody is raised in mouse and you are using an anti-mouse secondary antibody.

Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

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Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As discussed over the phone, Ihave attached a questionnaire so that I can gather further information regarding the samples tested and the protocol used. OnceI havereceive the completed questionnaire,I will look at the protocol and see if there are any suggestionsI can make that may improve the results.

This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

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