Human IgG1 ELISA Kit (ab100548)

Reviews (1)Q&A (12)References (1)

Overview

  • Product name

    Human IgG1 ELISA Kit
    See all IgG1 kits

  • Detection method

    Colorimetric

  • Sample type

    Cell culture supernatant, Saliva, Urine, Serum, Plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    < 0.05 ng/ml

  • Range

    0.05 ng/ml - 40 ng/ml

  • Recovery

    84 %

    Sample specific recovery
    Sample type Average % Range
    Serum 87 75% - 97%
    Plasma 81.5 71% - 94%

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Human

  • Product overview

    Abcam’s IgG1 Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human IgG1 in serum and plasma.

    This assay employs an antibody specific for Human IgG1 coated on a 96-well plate. Standards and samples are pipetted into the wells and IgG1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human IgG1 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IgG1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    Optimization may be required with urine samples

  • Tested applications

    Suitable for: Sandwich ELISAmore details

  • Platform

    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab100548 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Sandwich ELISA - IgG1 Human ELISA Kit (ab100548)
    Sandwich ELISA - IgG1 Human ELISA Kit (ab100548)

    Human IgG1 measured in biological fluids showing quantity (pg) per mL of tested sample. Human serum and plasma were diluted 1e6 - 64e6 fold. Human saliva and urine were diluted 1e3-64e3 fold.

  • Typical standard curve
    Typical standard curve

    Representative standard curve using ab100548

Protocols

References

This product has been referenced in:

See 1 Publication for this product

Publishing research using ab100548? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Detection of Trastuzumab in mouse plasma samples

 Good Average 3/5 (Ease of Use)
Abreviews
abreview image
The assay was tested for quantification of Trastuzumab plasma levels in a pharmacokinetic experiment in mice.
The standard curve was generated with both the recombinant IgG1 standard provided in the kit and the Trastuzumab, as comparison of relative O.D.
Each standard was assayed in duplicate, following the manufacturer instructions.
Plasma samples were diluted 10,000 fold with 1X Assay Diluent and gave the expected O.D. values.

Dr. Gianfranco Pasut

Verified customer

Submitted Apr 12 2017

Question

I understand that the capture antibody for kit Ab100548 is mouse monoclonal. We would like to use this kit to detect levels of human IgG1 in rat samples. Will the kit work, or would you expect cross reaction of the mouse capture antibody with rat IgG? Thank you.

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Answer

Hello! The laboratory is unsure if this would cross-react with rat or not, they have not tested it and have no data on this. Please let me know if you have any further questions.

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Question

What is the species that the polyclonal antibody is raised in?

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Answer

Thank you for contacting us.

Regarding our Human IgG1 ELISA kit, the host species for the capture antibody is mouse and it is monoclonal; the detection antibody is goat, polyclonal. I have attached the manual for your reference.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

When doing the final analysis, do you have to take the correction factor into account?

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Answer

Thank you for calling Abcam last week.

I have talked tothe lab and yes youwill need to multiplyyour sample concentration results by the sample dilution used in order to determine the actual levels of IgG1 in their samples. If it helps, the levels we measured testing normal serum with this kit was ˜7 mg/ml.

Please letme knowif you haveany questions or comments.

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Question

Our customer is going to purchase ab100548 and would like to know if there is a cross-reactivity with rat IgG.

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Answer

Cross-reactivity for ab100548 with rat has not been tested.
I can confirm that the capture antibody is monoclonal and the detection antibody is polyclonal. For the most part we do not see cross-species reactivity with sandwich based assays.

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Question

As promised I took a picture of the fungus growing in Item E. I received
the item E replacement from you and the new kit, thank you.
As well, I would like to thank you for the sweets, it was a nice thought.
In the future, I would suggest putting a label on item E to store at -20
(I have done that with the new batch you sent as well as the new kit since
that is allowed according to the data sheet). Or maybe if the supplier is
willing to filter the solution, it could also be a way to deal with this
problem.
In any case, thank you again for all your help and let me know if you
have any more questions.

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Answer

Thank you for your email and the image.

I will send it to the supplying lab. Also, thank you for your suggestions, which I will pass on as well. I can assure you that this problem with the item E has been in discussion recently and we are looking into solutions. Your suggestions will be of help.

I wish you good luck with your research and please let me know if I can help you with anything else.

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Question

Thank you for the info.
I have absolutely used ddh2o for the reconstitution of the item E. However,
that should not even
be an issue because the fungus also grew in the item E stock solution
which was in the fridge. I am currently away
for a conference, but I will take a photo of it in about 2 weeks when I
get back to the lab.

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Answer

Thank you for your reply.

Yes, you are making a good point there - the fungus growing in the stock solution without adding any water. I will let the lab know and proceed with arranging the replacement for item E.

Enjoy your conference and I will keep you updated. Yes, please send the image when you get back.
Thank you for your understanding.

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Question

ab100548: item E (assay diluent) has fungal contamination
after filtration obtained different results as before filtration of fungus
at the beginning: hazy solution, overtime, fungus grew bigger

lot GR65263-3 (Jan 2011) - used
lot GR71712-1 (Mar2012) - not used yet, but sees something growing in item E

ab100692: is plate coated with capture antibody only, or was protein A or G coupled to the capture antibody used for coating?

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Answer

Thank you for your patience.

I have heard back from the supplying lab with the following information:

For ab100692: The plate is only coated with the capture antibody.

For ab100548: Assay Diluent B does contain some protein and solutes so it would not be uncommon to see a little cloudiness or floating unsolubilized protein aggregates (especially after storing at low temperatures and/or for a long period of time). Most of the time bringing the solution up to room temperature, mixing it thoroughly and diluting to the 1X working concentration will help clarify the solution and solubilize any protein aggregates or precipitates. Centrifuging and filtering the solution can also help but usually doing so is not needed.

If this is whatthe customer is referring to, then they shouldn’t be concerned. However, if stored improperly and/or prepared using contaminated water (instead of distilled or deionzied water), fungal growth could occur.


I am almost certain that you used distilled or deionzied water, but would you mind confirming that?
Also, the lab would be interested to know ifyou happen tohave a photo of the diluent?

I have requested a vial of item E and am in discussion with them about that.

Please accept my sincere apologies about this delay in resolving the problem.

I look forward to hear back from you.

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Question

ab100548: item E (assay diluent) has fungal contamination
after filtration obtained different results as before filtration of fungus
at the beginning: hazy solution, overtime, fungus grew bigger

lot GR65263-3 (Jan 2011) - used
lot GR71712-1 (Mar2012) - not used yet, but sees something growing in item E

ab100692: is plate coated with capture antibody only, or was protein A or G coupled to the capture antibody used for coating?

Read More
Answer

Thank you for contacting us.

I am so sorry to hear about the problem you are having with our kits.

For ab100548, I have arranged for the replacement of 1 kit and am in touch with the lab to replace item E for the second kit.

As for ab100692, I have contacted the lab for more information and will be in touch as soon as possible.

In the meantime, could you please let me know your shipping address?

I look forward to hear back and to assist you in resolving this case.

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Question

Does this kit recognize the Fc or Fab fragment of the human IgG?

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Answer

Thank you for contacting Abcam

The capture antibody in the ab100548 kit recognizes the Fc portion of IgG1. The detection antibody recognizes the Fab portion of IgG. When used as a pair, the kit detects IgG1.

Please let me know if there is anything else I can help you with.

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1-10 of 13 Abreviews or Q&A

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