Mouse monoclonal [HP6025] Anti-Human IgG4 Fc (HRP) (ab99823)

Overview

  • Product name
    Mouse monoclonal [HP6025] Anti-Human IgG4 Fc (HRP)
    See all IgG4 secondary antibodies
  • Host species
    Mouse
  • Target species
    Human
  • Specificity
    ab99823 reacts with the Fc portion of the heavy chain of Human IgG4 as demonstrated by ELISA.
  • Tested applications
    Suitable for: ELISA, WB, Dot blot, ICC, IHC-Frmore details
  • Conjugation
    HRP

Properties

Applications

Our Abpromise guarantee covers the use of ab99823 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.
WB Use at an assay dependent dilution.
Dot blot Use at an assay dependent dilution.
ICC Use at an assay dependent dilution.
IHC-Fr 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) »
  • Lagatie O  et al. Evaluation of the Diagnostic Performance of Onchocerca volvulus Linear Epitopes in a Peptide Enzyme-Linked Immunosorbent Assay. Am J Trop Med Hyg 98:779-785 (2018). ELISA . Read more (PubMed: 29313477) »

See all 3 Publications for this product

Customer reviews and Q&As

Application
Other
To demonstrate the direct neutralizing effect of human anti-GLA IgG4 isotype antibodies, antibody-based α-Galactosidase A (GLA) enzymatic activity rescue assays were performed as follows. Fifteen µg of specific anti-IgG4 isotype antibody (mouse α-human IgG isotype 4 [ab99823] Abcam) were immobilized on a 96-well Maxisorp ELISA plate (Thermo Scientific) over night at 4°C. After 5 washes with 200 µl PBS/Tween 0.1%, blocking with PBS/BSA 2% (2 h at room temperature), 5 µg of purified total IgGs from individual patients (n=5 ERT-naïve as well as under treatment) with known inhibition status were applied in 100 µl PBS/BSA 2% (overnight at 4°C). Samples were washed trice with BPS/Tween 0.1%, blocked for 2 h at RT with PBS/BSA 2%, and incubated with 1 ng agalsidase-alfa in 50 µl PBS per well for 20 min. Subsequently, enzyme activity measurements were performed within the same wells by adding 50 µl reaction mix containing 4-methylumbelliferyl-α-D-galactopyranoside (Santa Cruz Biotechnology) as described elsewhere. After 1 h at 37 °C, supernatants (100 µl) were transferred into a new 96-well microplate, the reactions were stopped with 100 µl 0.5M Na2CO3 (pH 10.8) and GLA activities were measured. To determine rescued GLA activities, individual samples were normalized against BSA-treated GLA positive controls (BSA-treated).
Username

Abcam user community

Verified customer

Submitted Mar 02 2018

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]). 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.
Username

Abcam user community

Verified customer

Submitted Mar 02 2018

Application
Western blot
100 ng of agalsidase-alfa and -beta were blotted onto a PVDF membrane. Purified total IgGs from patients with Fabry disease were used as primary antibodies (end concentration in PBS/BSA 2%: 5.5 µg/ml). The discrimination between IgG1 and IgG4 isotypes was performed by using either mouse α-human IgG isotype 1 [ab99774] or mouse α-human IgG isotype 4 [ab99823] as secondary antibodies in a dilution according to manufacturer’s (Abcam) instructions. Anti-human IgG-HRP (sc-2907, Santa Cruz Biotechnology final concentration: 0.04 µg/ml) served as positive control.
Username

Abcam user community

Verified customer

Submitted Feb 27 2018

Thank you for contacting us.


We do know that ab99823 will bind to the Fc region of Human IgG4. We also offer ab99817 Mouse Anti-Human IgG4 (pFc')-HRP (Clone HP6023), which will bind to the pFc’ region (further away from the hinge re...

Read More

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 s...

Read More

Thank you for confirming those details.

I can suggest the goat anti-mouse antibodyhttp://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-H-L-HRP-pre-adsorbed-ab98808.htmlor the rabbit anti-mouse antibodyhttp://www.abcam.com/Rabb...

Read More

Thank you for contacting us and your interest in our products.

I am sorry but I have not completely understood the question. If you are asking if ab99823 can be used as a secondary antibody to ab67592 and ab52587 then the answer would be no. ...

Read More

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

Sign up