Overview

  • Product name

    Mouse IGFBP3 ELISA Kit
    See all IGFBP3 kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 70 pg/ml
  • Range

    54.87 pg/ml - 40000 pg/ml
  • Recovery

    89 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 105.5 93% - 112%
    Serum 85.83 73% - 94%
    Plasma 76.81 71% - 84%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Abcam’s IGFBP3 Mouse ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Mouse IGFBP3 in serum, plasma and cell culture supernatants.

    This assay employs an antibody specific for mouse IGFBP3 coated on a 96- well plate. Standards and samples are pipetted into the wells and IGFBP3 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse IGFBP3 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 IGFBP3 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • 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 C 1 x 30ml
    Biotinylated anti-Mouse IGFBP3 (lyophilized) 2 vials
    IGFBP3 Microplate (12 x 8 wells) 1 unit
    Recombinant Mouse IGFBP3 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

  • Relevance

    Insulin like growth factor binding protein 3 (IGFBP3) is a member of the superfamily of insulin like growth factor (IGF) binding proteins which include six high affinity IGF binding proteins (IGFBP) and at least four low affinity binding proteins referred to as IGFBP related proteins (IGFBPrP). The IGFBP members are cysteine rich proteins with conserved cysteine residues clustered in the amino terminal and the carboxy terminal regions of the molecule. The cDNA sequence encoding the mature human IGFBP3 is fused to the signal peptide of CD33. Human IGFBP3 is the major IGF binding protein in plasma where it exists in a ternary complex with IGFI or IGFII and an acid labile subunit. IGFBPs hold a central position in IGF ligand-receptor interactions through influences on both the bioavailability and distribution of IGFs in the extracellular environment. Insulin like growth factor binding protein 3 (IGFBP3) can modulate the mitogenic and metabolic effects of the insulin like growth factors (IGFs). Insulin like growth factor binding protein 3 (IGFBP3) is expressed in multiple tissues. The highest expression level is found in the non paranchymal cells of the liver. The expression levels are higher during extrauterine life and peak during puberty. Insulin like growth factors (IGFs) and IGF binding proteins (IGFBPs) play important roles in cell growth and differentiation. IGFBP3 is one of the factors in serum that is responsible for high serum induced apoptosis in PC3 cells, a prostate cancer cell line. IGFBP3 is important in controlling glucose homeostasis with increased urinary levels in type I diabetes with persistent microalbuminuria.
  • Cellular localization

    Secreted
  • Alternative names

    • Acid stable subunit of the 140 K IGF complex
    • Binding protein 29
    • Binding protein 53
    • BP 53
    • BP53
    • Growth hormone dependent binding protein
    • IBP 3
    • IBP3
    • IGF binding protein 3
    • IGFBP 3
    • Insulin Like Growth Factor Binding Protein 3
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab100692 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

  • IGFBP3 in mouse tissue lysates extracted with RIPA buffer. Data from kidney lysates diluted 1/50-1/250 (protein concentration 0.25-0.05 mg x mL-1) and lung diluted 1/10 (0.75 mg x mL-1) (duplicates +/- SD).

  • IGFBP3 in mouse biological fluids (duplicates +/- SD). Plasma and serum tested in the dilution range of 1/50-1/250, urine tested undiluted, rat and human samples (serum and plasma) were below level of detection at 1/10 dilution.

  • Representative Standard Curve using ab100692

Protocols

References

This product has been referenced in:

  • Suman S  et al. Space radiation exposure persistently increased leptin and IGF1 in serum and activated leptin-IGF1 signaling axis in mouse intestine. Sci Rep 6:31853 (2016). Read more (PubMed: 27558773) »
  • Suman S  et al. Protracted upregulation of leptin and IGF1 is associated with activation of PI3K/Akt and JAK2 pathway in mouse intestine after ionizing radiation exposure. Int J Biol Sci 11:274-83 (2015). Read more (PubMed: 25678846) »
See all 2 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Question
Answer

Thank you for contacting us at Abcam Scientific Support,

In regard to your enquiry on our IGFBP3 ELISA pair, we have not tested this interaction so we will not have the data to share which forms of IGFBP3 are available for binding by the assay. The free form is obviously recognized, while the bound for may or may not be recognized. As we do not map the location of the antibody epitopes we do not know yet what percentage, if any, of the bound form can be recognized. I hope this information is useful. Please let me know if I can provide any other help or technical advice.

Read More

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.

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.

Read More

Answer

Thank you for your inquiry.  I have addressed your questions below. Regarding the plate reader, any normal ELISA reader with colormetric detection which can read at 450nm and hold a standard 96 well plate should work.   The instructions for calculating the results are as follows: Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points. If you do not have ELISA analysis software like Sigma Plot you can use any other graphing software which can generate log-log, linear, or 4-parameter curves or even log-log graphing paper.  FYI, Excel is a good program to use for linear plots. Unfortunately, I was not able to find a reference for this particular kit.    I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.  

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Question
Answer

Thank you for contacting us. Yes, I can confirm that the Assay Diluent C is the same in both kits (ab100692, ab100693).   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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