Overview

  • Product name
    Human Insulin ELISA Kit
    See all Insulin kits
  • Detection method
    Colorimetric
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 4 µlU/ml
  • Range
    4.69 µlU/ml - 300 µlU/ml
  • Recovery

    89 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 76.16 68% - 88%
    Serum 91.4 83% - 102%
    Plasma 99.03 73% - 128%

  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Cow
  • Product overview

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


    This assay employs an antibody specific for Human Insulin coated on a 96-well plate. Standards and samples are pipetted into the wells and Insulin present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human Insulin 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 Insulin bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    This kit detects both insulin and proinsulin.


    Get higher sensitivity in only 90 minutes with Human Insulin ELISA Kit (ab200011) from our SimpleStep ELISA® range.

  • Notes

    Optimization may be required with urine samples.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    500X HRP-Streptavidin Concentrate 1 x 200µl
    5X Assay Diluent B 1 x 15ml
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human Insulin (lyophilized) 2 vials
    Insulin Microplate (12 strips x 8 wells) 1 unit
    Recombinant Human Insulin Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas
  • Function
    Insulin decreases blood glucose concentration. It increases cell permeability to monosaccharides, amino acids and fatty acids. It accelerates glycolysis, the pentose phosphate cycle, and glycogen synthesis in liver.
  • Involvement in disease
    Defects in INS are the cause of familial hyperproinsulinemia (FHPRI) [MIM:176730].
    Defects in INS are a cause of diabetes mellitus insulin-dependent type 2 (IDDM2) [MIM:125852]. IDDM2 is a multifactorial disorder of glucose homeostasis that is characterized by susceptibility to ketoacidosis in the absence of insulin therapy. Clinical fetaures are polydipsia, polyphagia and polyuria which result from hyperglycemia-induced osmotic diuresis and secondary thirst. These derangements result in long-term complications that affect the eyes, kidneys, nerves, and blood vessels.
    Defects in INS are a cause of diabetes mellitus permanent neonatal (PNDM) [MIM:606176]. PNDM is a rare form of diabetes distinct from childhood-onset autoimmune diabetes mellitus type 1. It is characterized by insulin-requiring hyperglycemia that is diagnosed within the first months of life. Permanent neonatal diabetes requires lifelong therapy.
    Defects in INS are a cause of maturity-onset diabetes of the young type 10 (MODY10) [MIM:613370]. MODY10 is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease.
  • Sequence similarities
    Belongs to the insulin family.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Alternative names
    • IDDM2
    • ILPR
    • ins
    • INS_HUMAN
    • Insulin
    • Insulin A chain
    • Insulin B chain
    • IRDN
    • MODY10
    • Preproinsulin
    • Proinsulin
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab100578 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

  • Human Insulin measured in cell culture supernatants showing quantity (µIU) per mL of tested sample. Samples diluted 1-2 fold.

  • Human Insulin measured in biological fluids showing quantity (µIU) per mL of tested sample. Samples diluted 1-6 fold.

  • Human Insulin standard curve: mean of duplicates (+/- SD), performed in Diluent A

  • Representative Standard Curve using ab100578.

  • Representative Standard Curve using ab100578.

Protocols

References

This product has been referenced in:
  • Citro A  et al. Biofabrication of a vascularized islet organ for type 1 diabetes. Biomaterials 199:40-51 (2019). Read more (PubMed: 30735895) »
  • Xu Y  et al. Effects of citrate ester plasticizers and bis (2-ethylhexyl) phthalate in the OECD 28-day repeated-dose toxicity test (OECD TG 407). Environ Res 172:675-683 (2019). Read more (PubMed: 30878739) »
See all 16 Publications for this product

Customer reviews and Q&As

1-10 of 33 Abreviews or Q&A

Answer

Thank you very much for your interest in ab100578. To our knowledge, ab100578 has not been tested in non-human primate. Therefore, I can offer a discount off a future purchase if you buy ab100578 now, test it in non-human primate and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free kit of the same value. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab100578 in non-human primate. I will then send a discount code. This code must be issued before purchasing ab100578 so please wait for my reply before ordering. 2. Purchase ab100578 either by phone, fax, or online (www.abcam.com). 3. Test it in non-human primate. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any kit ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab100578 turns out to be unsuitable for non-human primate you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.  

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Abreviews
We used the kit to detect insulin in normal WT mouse serum or plasma. We tested 1 to 25ul of serum or plasma per well but everything was BLQ. The standard curve however was beautiful so I believe this kit may work for other samples. In our case we switched to a more sensitive detection method.

Abcam user community

Verified customer

Submitted Oct 23 2018

Question
Answer




I am happy to confirm that Heparin, EDTA and Citrate treated plasma samples were all tested with this kit and are all suitable. This is covered by our AbPromise guarantee.


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Application to plasma samples from pigs

Good Excellent 5/5 (Ease of Use)
Abreviews
Plasma samples from pigs 3-4 months old (25-30 kg body weight) were obtained after overnight fasting.

Assay procedure was performed according to manufacturer instructions. However, standard curve (duplicates) was modified due to the low concentrations observed in fasted pigs. Standard concentrations were 100, 50, 25, 12.5, 6.25, 3.125 and 1.5625 microIU/mL, prepared by a 1:3 dilution of the first standard provided with the product, and consecutive 1:2 dilutions thereafter.

Plasma samples were added to each well by duplicate (100 microL) as suggested by the manufacturer instructions, with no dilution. All remaining steps were performed in accordance with manufacturer instrucitions, with no modificatons. Readings were performed at 450 nm.

Results were as follows:
Control animals: 6.63, 3.65, 1.10 and 4.67 microIU/mL (mean: 4.01±1.15 microIU/mL)
Treatment A: 54.95, 9.29, 8.77, 5.10 microIU/mL (mean: 19.53±11.84).

Mr. Antonio Rodriguez Sinovas

Verified customer

Submitted Oct 28 2014

Answer

We do not have data for camel samples tested with the human insulin ELISA, ab100578. However, insulin is well-conserved between the two species, so it is possible the ELISA will detect camel insulin.

To check the similarity between the two species, I used the camel UniProt database entry at the following link:

P01320 (INS_CAMDR) http://www.uniprot.org/uniprot/P01320

The pig insulin ELISA ab156515 will be a slightly better choice, since camel insulin is more similar to pig insulin than human.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=156515).

Here are the percentage identity results of the alignments of camel insulin precursor with human and pig insulin.

93% w/ human insulin B chain http://www.uniprot.org/blast/?about=P01308[25-54]
90% w/ human insulin Achain http://www.uniprot.org/blast/?about=P01308[90-110]

97% w/ pig insulin B chain http://www.uniprot.org/blast/?about=P01315[25-54]
90% w/ pig insulin A chain http://www.uniprot.org/blast/?about=P01308[90-110]

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Question
Answer

Thanks for your enquiry.

The kit can be used within 1 year if the entire kit is stored unopened at -20 degrees C. After opening, please refer to the protocol booklet for component-specific storage instructions.

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Answer

The general sample information for the Insulin Human ELISA Kit (ab100578) is a suggested dilution of normal serum/plasma: 2-10 fold (page 10 of the Protocol Booklet).
We however recommend testing different dilutions such as 1:2, 1:5, 1:10 to determine which one is the best for your particular sample.
Once the sample has been diluted, 100ul of sample should be added into appropriate wells and incubate for 2.5 hours at room temperature or over night at 4C with gentle shaking (remember to cover well the plate). For the following steps please follow the kit protocol at page 11 of the Protocol Booklet.

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Abreviews
This kit initially intended to detect human Insulin, successfully detect also rat Insulin in serum samples.

Abcam user community

Verified customer

Submitted Jul 05 2013

Question
Answer

The antibodies used in this kit will recognize Insulin and Proinsulin. From the information which I have been able to gather on SwissProt regarding insulin (http://www.uniprot.org/uniprot/P01308) the insulin precuror is composed of a signal peptide, chains A, B and a C peptide. It seems that insulin is the heterodimer of chain A and chain B and that proinsulin contains chain A, B and peptide C. While I cannot be sure of whether the proinsulin is non functional it seems likely that this form could be compared to the functional heterodimer. If this is incorrect please let me know. Assuming that this is correct, then ab100578 will recognize both forms. I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

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Question
Answer



Les anticorps utilisés dans le kit ELISA ab100578 sont les anticorps ab8304 comme anticorps de capture et ab20756 comme anticorps de détection. Ces deux anticorps sont testés et garantis chez le rat. Le kit lui-même n’a pas été testé chez le rat mais il y’a des chances que cela fonctionne. Si vous souhaitez le tester, je peux vous faire une offre de test.

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1-10 of 33 Abreviews or Q&A

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