Mouse Leptin ELISA Kit (ab100718)

We have not yet tested ab100718 with cell lysates. However, the kit is compatible with many different samples types so lysates will most likely work. If you decide to test this kit with lysate samples, we recommend diluting the samples at least 5-fold with Assay Diluent B to minimize any effects of the detergents in the lysis buffer. The samples may need to be diluted further but this would need to be determined by you empirically. In brief, a lysis buffer must meet the following specifications: A) has relatively low salt content (700 mM or less) B) does not contain sodium azide C) does not contain >0.1% SDS D) does not contain >10 mM reducing agents (beta-mercaptoethanol or dithiothreitol).

Thank you very much for your inquiry and your interest in our kits.

Unfortunately,the cross-reactivity of ab108879and ab108881 with pig samples is low, around 5%.

For the other three kits, ab100581, ab100718 and ab100773,the laboratory has not tested these on pig samples and no prediction can be made, as not all epitopes are known.

For the most part we do not see cross-species reactivity with sandwich based assays. If you would however nevertheless like to test on of these kits, please do let me know, as I would be able to provide you with an interesting testing offer.

Please do also let me know whether you would like to discuss other options. Indeed, we have many individual antibodies against leptin in our catalog as well. Some of them are tested in pig (ab3583 and ab16227) and others are tested in ELISA. Again, we could discuss options and provide testing offers if you are interested in this.

The testing offers would be along the line of the offer described here:
https://www.abcam.com/collaborationdiscount

I hope this information is helpful. I am looking forward to hear back from you.

Thank you for your enquiry.

I have now received further information regarding the reading of the results from this kit. The manuals advise using a log-log model as this type of plot will fit most data sets very well. However, you can use any graphing model including linear regression, 4-parameter, 2-order polynomial, etc.When testing this kit, theoriginator ofthe kit plots theELISA data using log-log, linear regression, AND 4-parameter and then choose the model that fits the data best.

I hope this is helpful to you. If you have any further questions, please do not hesitate to contact us.

Thank you for your inquiry.  This kit ab100718 has been validated for serum, plasma, and cell culture supernatant samples. Are you using whole blood samples?  If so, this kit may work, but we would not recommend it as the chances of experiencing high background and interference would increase.  Below are some tips on sample preparation: How do I make preparation of blood and serum samples? For plasma: Collect blood into EDTA or Sodium heparin tube (e.g. BD vacutainer, Cat. No: 8001302 or 16852) Spin 10 minutes at 3000 rpm Aliquot into small tubes and store at –800C until use. For serum: Collect blood into tube without additive (e.g. BD vacutainer, Cat. No. 8002527) Keep at room temperature for 20 minutes Spin 10 minutes at 3000 rpm Aliquot into small tubes and store at –800C until use. Our recommended mouse serum/plasma sample dilution for ab100718 is ~10-40X.  Please note that this is just a recommendation and that the optimal dilution (if any) should be determined by the end-user empirically based on researched literature and knowledge of their particular samples.  Finally, 200ul of working volume per sample (100ul per well) is required if running each sample in duplicate (advised).  Additional information on the reagents and protocol can be found in the protocol booklet. https://www.abcam.com/ps/products/100/ab100718/documents/Leptin-Mouse-ELISA-Kit-ab100718-protocol-plain.pdf I hope this information helps. Please contact us with any other questions.