Mouse monoclonal [4E3] Anti-Human IgG1 hinge heavy chain (HRP) (ab99774)

Overview

  • Product name

    Mouse monoclonal [4E3] Anti-Human IgG1 hinge heavy chain (HRP)
    See all IgG1 secondary antibodies
  • Host species

    Mouse
  • Target species

    Human
  • Tested applications

    Suitable for: ELISA, ICCmore details
  • Immunogen

    Other Immunogen Type corresponding to IgG1.

  • Conjugation

    HRP

Properties

Applications

Our Abpromise guarantee covers the use of ab99774 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
ELISA 1/1000 - 1/2000.
ICC Use at an assay dependent dilution.

References

This product has been referenced in:

  • Lenders M  et al. Characterization of drug-neutralizing antibodies in patients with Fabry disease during infusion. J Allergy Clin Immunol N/A:N/A (2018). Read more (PubMed: 29421273) »
  • Perugino CA  et al. Identification of galectin-3 as an autoantigen in patients with IgG4-related disease. J Allergy Clin Immunol N/A:N/A (2018). ELISA . Read more (PubMed: 29852256) »
See all 5 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
ELISA
To determine the IgG isotype of neutralizing anti-drug antibodies, ELISAs were performed as follows. 96-well Maxisorp ELISA plates (Thermo Scientific) were coated overnight at 4°C with 50 ng agalsidase-alfa, washed 3x with PBS, and blocked for 2h at RT with 100 µl PBS/BSA 2% (per well). Subsequently, wells were washed 5x with PBS/Tween 0.1%. Purified total IgGs from patients) were used as primary antibodies and were applied in serial dilutions (1:250 1:500 1:1,000) in PBS/BSA 2% overnight at 4°C. Next day, cells were washed again 5x PBS/Tween 0.1% and incubated for 1 h at RT with IgG isotype-specific anti-human IgG-HRP (mouse α-human IgG isotype 1 [ab99774] mouse α-human IgG isotype 2 [ab99779] rabbit α-human IgG isotype 3 [ab86253] mouse α-human IgG isotype 4 [ab99823] goat α-human IgA [ab97215] mouse α-human IgE [ab99806]). Rabbit α-human IgG isotype 3 was detected with goat α-rabbit IgG-HRP [sc-2054, Santa Cruz Biotechnology,] as secondary antibody (diluted 1:5,000).
After 5 additional washes, 100 µl 1-Step Ultra TMB-ELISA Substrate Solution (Thermo Scientific) was added to each well, incubated for 10 min, and stopped with 2M sulfuric acid. The absorption was measured at 450 nm and ratios were calculated between Enzyme replacment therapy-naïve and -treated status.

Abcam user community

Verified customer

Submitted Feb 27 2018

Answer

Thank you for your inquiry.

The concentration is lot specific of these antibodies and will not be written on the bottle. I am however very happy to check this for you - I am currently checking the concentrations of the lots we are having in stock.

If you have not yet received the antibodies, I can check the concentrations of the current lot. If you have already ordered the antibodies, please provide me with the lot numbers of the antibodies you have, and I will check the concentration with the laboratory for the respective antibodies.

If you do not have the lot numbers, please provide me with the order information, so I can trace back the lot numbers myself.

I look forward to hear back from you.

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Answer

Following on my previous message about the secondary antibodies you are interested in, the anti-human IgG isotypes are tested on a Human IgG subclass plate (IgG1, IgG2, IgG3, and IgG4) to ensure activity and minimal cross reactivity with the other IgG subclasses.

mouse mAb anti human IgG1-HRP #ab99774

mouse mAb anti human IgG2-HRP #ab99784

mouse mAb anti human IgG3-HRP #ab99829

mouse mAb anti human IgG4-HRP #ab99817

Please let me know if you need the data.

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