Human GM-CSF ELISA Kit (ab100529)

Submit a review Q&A (12)References (1)

Overview

  • Product name

    Human GM-CSF ELISA Kit
    See all GM-CSF kits

  • Detection method

    Colorimetric

  • Sample type

    Cell culture supernatant, Serum, Plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    < 2 pg/ml

  • Range

    2.05 pg/ml - 500 pg/ml

  • Recovery

    > 75 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 101.1 89% - 114%
    Serum 75.43 69% - 83%
    Plasma 74.49 68% - 83%

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Human

  • Product overview

    Abcam’s GM-CSF Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human GM-CSF in serum, plasma, and cell culture supernatants

    This assay employs an antibody specific for Human GM-CSF coated on a 96-well plate. Standards and samples are pipetted into the wells and GM-CSF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human GM-CSF 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 GM-CSF 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 ab100529 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

  • Typical standard curve
    Typical standard curve

    Representative standard curve using ab100529

Protocols

References

This product has been referenced in:

  • Yang M  et al. Chemotherapy induces tumor immune evasion by upregulation of programmed cell death ligand 1 expression in bone marrow stromal cells. Mol Oncol 11:358-372 (2017). Read more (PubMed: 28218497) »
See 1 Publication for this product

Publishing research using ab100529? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 12 Abreviews or Q&A

Question

Product code: 100529
Lot number: GR86435-1
Inquiry: Dear Sir, with attachment the results (OD of standered curve) of my work on GMCSF for 2 kit , please let me know if these results are good so I can evaluate the results of my patients which I had it. Thanks and Best Regards

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Answer

Thank you for contacting us.

I have reviewed the data you have kindly sent and I am assuming that the data is for standard curve. The readings seems not good enough because the OD for 0 is comparable to 2.05pg/ml. The OD for "0" pg/ml actually gives the background reading which should be subtracted from the other readings of standard and the test sample.

These readings from whole standard curve also not show an increasing pattern with standard concentration until 32pg/ml which is actually lower than 5.12pg/ml well.

My suggestions would be repeating the experiment by carefully
- avoiding any pipetting error
- washing the plate
- making workable dilutions for standard, HRP-Streptavidin and detection antibody.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

They are free ! What we should pay??????

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Answer

Yes, It is all free however they need to check if the items were paid in original order.

It is just a normal check so there is no need to worry, you will be getting a order confirmation soon.

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Question

I didn't hear about the order shipping .. etc.. Please let me know ASAP, as the customer waiting !
Truly yours,

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Answer

Our accountsdepartment is holding the order at the moment. The order will be shipped once they authorize it, after checking thepayment etc. Please email mailto:creditcontrol@abcam.com for any order update.

Thanks!

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Question

Please see below:

thank you for your reply,

I have worked the tow kits on the same day as the incubation time between the kits is enough and when I got a step addition of HRP-Streptavidin noted deficiencies and tried to use the second kit to avoid the lack of the first I noticed the same problem, so i don't have any kit.
please I need the kits urgently.

Waiting your reply !

Truly yours,

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Answer

Thank you for contacting us.

To prevent the shortageissue in the future, please advise your customer along with other customers to check all reagents before beginning the experiment and that if there is a volume shortage or any other issue to not try and use this reagents without first contacting us to prevent what is most of the time an easily correctable and avoidable problem.

We have place order for 2 kits ab100562that will be shipped to you soon. The order number is 1130294.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

thank you for your reply,I have worked the tow kits on the same day as the incubation time between the kits is enough and when I got a step addition of HRP-Streptavidin noted deficiencies and tried to use the second kit to avoid the lack of the first I noticed the same problem, so i don't have any kit.
please I need the kits urgently.
"
Waiting your reply !
Truly yours,

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Answer

Thank you for your cooperation.

I have sent a request to lab for new kits and hoping to get an answer later this evening. I will confirm you the dispatch date.

I hope this info will be helpful.

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Question

Thanks a lot for your support, it is really appreciated !
Please see below reply from the end user:
Thank you and thanks Abcam for your help and support, kindly remember it was 2 kits of ab100562 cause I have 176 sample to run, I will hoping to receive the kits soon .
As for reagentHRP-streptavidin in ab100529 the amount was enough but the standared for the 2 kits of GM-CSF were in efficient which gave me bad standered curve and unclear result.

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Answer

Thank you for your email.

Regarding ab100562 have you used both kits? If not, then I will send you one replacement kit and one extra vial of HRP-standard. Could you let me know if you still have to use one kit of ab100562?

There are many factor that affect the standard curve. Could you please check the following when doing experiment;

- the reagents should be at room temperature when used
- avoid any pipetting error
- keep TMB substrate away from light
- check the ELISA reader wavelengths
- check standard is properly reconstituted and mixed
- try incubating the primary antibody for 24 hours.

I am sure these tips can help you get good results. Please do not hesitate to contact me if results does not improve.

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Question

Thank you for your prompt reply.

Concerning stock item G, i centrifuged the vial before taking the liquid out, after spinning i recovered only 10µl. The failure of the experiment i expect was dueto the insufficient amount ofHRP-strepatividin provided in the kit.What i did is i diluted the 10µlof stock item G into 10 ml diluent in order for it to be enough for the entire 96 well plate to avoid the error that happened, therefore i did not get any good results for the entire plate.

thanks for your attention.

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Answer

That's fine!

I will be sending you a replacement kitofab100562.

Could you check the HRP-streptavidin amount in ab100529.Is it OK?

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Question

Kindly be informed that you have supplied me recently with two kits ofInterleukin 1 beta Human ELISA (code: ab100562)(lot:GR85228-1)from Abcam Company which were unable to give me any result because of the following:
- The kit should contain 200µl of 300x concentrated HRP-conjugated streptavidin (item G in kit); whereas ifound less than 10 µl in the vial of each kit, knowing that i need (at least) 40 µl to prepare enough solution for 96 wells according to step 7 in the abcam leaflet,therefore due to the lack of this itemthe work failed and i didn't gain any results.
I also received two other kits that are GM-CSF Human Elisa kit (code:ab100529)(lot:GR86435-1), concerning these two kits i kindly need the normal range and the typical assay data for standard curve for kit in order tocompare them with my results.
Appreciate your cooperation and support.
Thanks and best regards,

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Answer

Thank you for your email. I apologize for any inconvenience caused.

The volume 200ul is actually more than what is need for the assay; this extra volume is added to avoid any loss due to evaporation, insufficient centrifuging, etc. Actually only 32ul X 300ul = 9.6 ml is needed for full 96 well assay plate.

I have personally checked the in-stock Item G of same lot which you have received. The volume was ˜50 ul, I accept it is not 200ul so I have informed our lab to tighten the caps of all vials. 10ul as you mentioned is quite less so I would like to ask, if you spin the vial in centrifuge before taking the liquid out? We always recommend our customers spinning the vials, in order to recover maximum amount possible.

Could you tell us more about the problem? Was your experiment failed due to insufficient HRP-strepatividin or due to other factors. Did you get standard curve? The recovered 10ul solution would have been enough for 25-30 wells; did you not get good results?

The kit ab100529 can detect GM-CSF in range between 2-500 picogramm/ml.

The standard should be made by the end user. Please check the protocol booklet for details about standard curve and detection range.

I hope this information will be helpful. I will be looking forward to hearing from you soon.

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Question

Dear sir,

U told me the range for the product



Product code: 100529, Lot number: GR86435-1 is
2.05 pg/ml - 500 pg/ml ,Is this the normal value if not please provide me with the normal value according to age and gender (to determain the abnormal result from healthy result),also i need the typical assay data for standard curve.

Thanks and best regards

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Answer

Thank you for your reply.
I confirm that 2.05 pg/ml - 500 pg/ml is the range the kit detects.
Please consult the appropriate literature for the normal concentration for typical concentration of GM-CSF in healthy humans.
A typical standard curve can be found in the protocol booklet. These standard curves are for demonstration only. A standard curve must be run with each assay.
I hope 5this information is helpful and wish you good luck with your experiments.

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Question

Product code: 100529
Lot number: GR86435-1

Inquiry: Good Day, Please i need the normal range for this prosuct to assase product result. thanks and best regards

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Answer

Thank you for your inquiry.
I am happy to confirm that the range of this kit is typically 2.05 pg/ml - 500 pg/ml as stated on the datasheet.
We also have data on the recovery that can also be found on the datasheet and in the protocol booklet:
https://www.abcam.com/index.html?datasheet=100529 https://www.abcam.com/index.html?datasheet=100529.

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