Overview

  • Product name

    Anti-IGF1 antibody (Biotin)
    See all IGF1 primary antibodies
  • Description

    Rabbit polyclonal to IGF1 (Biotin)
  • Host species

    Rabbit
  • Conjugation

    Biotin
  • Tested applications

    Suitable for: Sandwich ELISA, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Highly pure (>98%) recombinant human IGF1.

Properties

Applications

Our Abpromise guarantee covers the use of ab83137 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 a concentration of 0.25 - 1 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to IGF1 (ab9572). Allows for the detection of at least 0.2 - 0.4 ng/well of recombinant IGF1.
WB Use a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant hIGF-I is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions.

Target

  • Function

    The insulin-like growth factors, isolated from plasma, are structurally and functionally related to insulin but have a much higher growth-promoting activity. May be a physiological regulator of [1-14C]-2-deoxy-D-glucose (2DG) transport and glycogen synthesis in osteoblasts. Stimulates glucose transport in rat bone-derived osteoblastic (PyMS) cells and is effective at much lower concentrations than insulin, not only regarding glycogen and DNA synthesis but also with regard to enhancing glucose uptake.
  • Involvement in disease

    Defects in IGF1 are the cause of insulin-like growth factor I deficiency (IGF1 deficiency) [MIM:608747]. IGF1 deficiency is an autosomal recessive disorder characterized by growth retardation, sensorineural deafness and mental retardation.
  • Sequence similarities

    Belongs to the insulin family.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Database links

  • Form

    There are 2 isoforms produced by alternative splicing. Isoform 1 also known as: IGF-IB; Isoform 2 also known as: IGF-IA.
  • Alternative names

    • IBP1 antibody
    • IGF I antibody
    • IGF IA antibody
    • IGF IB antibody
    • IGF-I antibody
    • Igf1 antibody
    • IGF1_HUMAN antibody
    • IGF1A antibody
    • IGFI antibody
    • IGFIA antibody
    • Insulin like growth factor 1 (somatomedin C) antibody
    • Insulin like growth factor 1 antibody
    • Insulin like growth factor IA antibody
    • Insulin like growth factor IB antibody
    • Insulin-like growth factor I antibody
    • Mechano growth factor antibody
    • MGF antibody
    • OTTHUMP00000195080 antibody
    • OTTHUMP00000195081 antibody
    • OTTHUMP00000195082 antibody
    • OTTHUMP00000195083 antibody
    • OTTHUMP00000195084 antibody
    • Somatomedia C antibody
    • Somatomedin C antibody
    • Somatomedin-C antibody
    see all

References

ab83137 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Answer

Thank you for your patience.

I try to help you as much as possible while my colleague is out of the office. I have read through all the e-mails related to your enquiry. In order to be able to identify the sources of the problem, it would be interesting to know more about the experiments:

1) Samples:

- Are you using exactly the same samples i.e. aliquotted the serum and stored in the freezer? Or the values represent the results of independent samples?

- Have you carried out a head-to head experiment with the ELISA kit and the Sandwich ELISA using the same sample?

- Are the samples from normal healthy individuals or from patients?

2) Free vs bound IGF1:

It would be interesting to know what the objectives of your studies ie. quantifying the free, or the bound or the total IGF1?

I look forward to hearing from you soon.

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Answer

Thank you for getting back to me. I had not as yet sent the information I have in regards to the antibodies and their use in ELISA as I was still waiting for some more, but I'll share what I have now.

The anti-IGF1 antibody (Biotin) (ab83137) has been used to perform sandwich ELISAs previously. This was done using the anti-IGF1 antibody (ab9572) as the capture antibody, and the protein ab9573 was used as the standard. The two antibodies, ab83137 and ab9572, are both rabbit polyclonals raised to human IGF1.

As mentioned previously, the Mouse monoclonal [1F6-3H10] to IGF1 (ab40789) has as yet only been used in indirect ELISA as the detection antibody. I have not as yet been able to identify exactly which secondary antibody was used in conjunction with this but the secondary antibody https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG1-heavy-chain-HRP-pre-adsorbed-ab98693.htmlshould be suitable for this purpose.

As we have no images currently for either ab83137 or ab40789, you would be eligible for our testing discount.This offer means that if you purchase ab83137 or ab40789 as normal, test the antibody with your ELISA and let us know of the results and an image through an Abreview (no matter whether positive or negative) you would be eligible for a free primary antibody of your choice from our catalogue (or the equivalent price of ab83137 or ab40789 off any product). More information on this offer can be found here:

www.abcam.com/collaborationdiscount

Please note that the antibody to be tested must be purchased, tested, the Abreview submitted and the free product claimed within a 4 month period.

If you would be interested in participating in this scheme please do let me know as a discount code needs to be issued prior to the purchase of ab83137 or ab40789.

I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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Answer

Thank you for getting back to me.

We do have one antibody which has been used in sandwich ELISA previously to detect human IGF1. This is antibody https://www.abcam.com/IGF1-antibody-Biotin-ab83137.htmland it is biotin conjugated. I am enquiring further as to which other antibody was used with this antibody as a partner. I will get back to you as soon as I have this information.

I am also seeking further information as to how ab40789 has been used in indirect ELISA and will again get back to you as soon as I have this information.

I am sorry for the delay and any inconvenience this may cause you.

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Answer

Thank you for your message and for providing this further information.

To confirm, I can confirm this antibody has been tested and therefore guaranteed for human samples. The previous email stated that it has not been tested specifically with serum samples in ELISA. Serum contains many other proteins and other biological factors that may be interfering with the antibody binding.

I am sorry to hear the suggestions made have not improved the results on this occasion. As previously discussed,the antibodies seem to be workingbut regrettably there seems to be some interference from something within the serum samples. I appreciate the time you have spent on these experiments and as I gesture of goodwill I would be pleased to arrange a credit note. Alternatively, Iwould be pleased to provide free of charge replacements if this is what you prefer and would like to try again. However, please bear in mind it is possible this may not solve the issue if there is a possibility the serum sample type is not suitable for use with these antibodies in ELISA.

I hope this will provide a satisfactory resolution. I look forward to hearing from you with details of how you would like to proceed.

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Question

Dear Kate,



Thank you for your email. As the antibodies are guaranteed to work with serum samples, what kind of pre-treatment would I need to use in order to get accurate results? What was used during testing for IGF1 ELISA?



Please find my replies to the questionnaire below in blue:







Order Details

Antibody code: ab9572 and ab83137



Problem

Choose: weak signal with serum samples but not with standard curve





Lot number GR21525-6 (ab9572) and GR9925-6 (ab83137)





Purchase order number HFE2462590-1 (ab83137) and ab9572 was sent for free as per the abpromise

or preferably Abcam order number





General Information

Antibody storage conditions (temperature/reconstitution etc)

Aliquoted ab9572 in 5ul aliquots and stored at -20C. Reconstituted ab83137 in PBS/0.1% BSA, aliquoted as 5ul aliquots and stored at -20C.



Description of the problem (high background, no signal, non-specific color development, poor standard curve etc.)



While the standard curve is good, the pre-treated serum samples are the problem. The serum is from samples that had already been tested using an IGF1 ELISA kit. This serum (from the same tube) was stored at -80C. For the current ELISA, I pre-treated the serum by first treating 30ul of serum with 120ul acid-ethanol (2.5% HCl-87.5% ethanol). This was then incubated at room temperature for 30 minutes with shaking. The samples were then centrifuged for 5 minutes at 10,000rpm and 100ul of supernatant was transferred to a fresh tube containing 200ul Tris Buffer (pH 7.6). The tube was mixed thoroughly and assayed immediately. I also assayed the same serum samples in a 1:15 dilution using PBS to check if the pre-treatment was working. Additionally, I had human serum control (from a bottle from Sigma) as an additional sample. So this was pre-treated and only diluted as well. However, the absorbance and the concentrations calculated from the standard curve for both the pre-treated samples and the diluted samples were similar and these concentrations were about 4-5-fold lower than previously obtained results for the same samples.







Type of ELISA (Direct ELISA/Indirect ELISA/Sandwich ELISA etc.)

Sandwich ELISA



Sample (Species/Cell type/Cell line etc.)

Serum samples either pre-treated or not.



Coating well (Buffer/Concentration of the coating material etc.)

ab9572 was used to coat the plates at 1:1000 dilution in PBS. The coated plates were stored at 4C overnight.



Blocking conditions (Buffer/time period, Blocking agent etc.)

5% non-fat milk in PBS solution was used to block the plates for 1 hour.



Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

ab83137 was used as the detection antibody at 1:6000. The plates were incubated at room temperature for 90 minutes. Wash steps were done 6 times in PBS/Tween 20.



Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

Streptavidin-HRP (Sigma) at 1:2000 dilution in PBS. Incubated for 90 minutes, wash 6x in PBS/Tween20.



Detection method (Substrate/Diluent etc.)

Citrate buffer with TMB and hydrogen peroxide.



Positive and negative controls used (please specify)

The serum samples (both pre-treated and diluted) were used as positive controls. I used two samples, one with a high concentration (˜400 ng/ml) and the other with a lower concentration (˜55 ng/ml). Also, blanks were used.







Optimization attempts (problem solving)

How many times have you tried the ELISA?

Two times with the samples; 5 times with standards and a human serum control (Sigma).





Have you run a "No Primary" control?

Yes

Do you obtain the same results every time?

The problem is not with the capture and detection antibodies not detecting IGF1 but with very low absorbance/signal for serum samples, either pre-treated or diluted.





Additional Notes:



Data:

We would appreciate if you are able to provide a copy of the data, particularly from the standard curve. This will help us to assess the results.



I have attached an Excel file with the results from the second test with samples.







Help us improve our service.

Rate your experience with us today. <https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3782221>



Best regards,

Kate



Kate Hayes

Scientific Support Supervisor

Abcam plc

https://www.abcam.com



Your original inquiry to Abcam:













Dear Kate,

Thank you for your reply. During testing for IGFBP3 and IGF1 antibodies, were the ELISAs tested with serum/plasma samples at all? Do you have any other customers who have used either sets of antibodies to set up a sandwich ELISA? I am having trouble with the IGF1 antibodies with pre-treated serum samples. I was wondering if you have any protocols for serum/plasma pre-treatment to use for this protocol?

Thank you,









Abcam Customer Services and Scientific Support Team

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Answer

Thank you for your reply

Please note that theseantibodies have not been tested with with serum samples, and so I am sorry no pretreatments have been tested which we could provide you with information on. In theory the antibodiesshould work with serum samples. Since the antibodies are working well with their standard protein, this means that the antibodies do work in the ELISA.Therefore, I can suggestit may be that there is some interferencewith something in the serum samples.

1. Have you been able to confirm the expected amount of protein in their samples using any other method?

2. Is it possible that the samples contain a very small amount of protein?

3. Is there any azide in the samples? This could inhibit the reactionand would lead to loss ofdetection as you are observing.

4. Have you tried any other sample types?

5. Please provide details of the sample preparation.

I hope this will be helpful. Please do not hesitate to get back to me with the requested information and I hope we can resolve this for you as soon as possible.

Read More

Answer

Thank you for your reply.

I would like to reassure you that ab83137 Anti-IGF1 antibody (Biotin) and ab9572 Anti-IGF1 antibody are tested and covered by our 6 month guarantee for sandwich ELISA together, and human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

As the difficulties are continuing, I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you could include details of the sample pretreatment. What was the pretreatment protocol, and the reasons why this was done?

I would appreciate if you could also providethe data from the standard curve and samples which would help us to assess the results.

With regards to the protocol used for testing, both the IGF1 and IGFBP3 antibodies will have been tested using the same procedure. Once I have further information from you, I would like to investigate further this further with our source if they have any other recommendations for you from their experiences with this antibody, particularly with regards to sample type and preparation.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


Order Details
Antibody code:

Problem
Choose: Problem with standard curve No signal or weak signal High background


Lot number


Purchase order number
or preferably Abcam order number


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, no signal, non-specific color development, poor standard curve etc.)


Type of ELISA (Direct ELISA/Indirect ELISA/Sandwich ELISA etc.)


Sample (Species/Cell type/Cell line etc.)


Coating well (Buffer/Concentration of the coating material etc.)


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (Substrate/Diluent etc.)


Positive and negative controls used (please specify)




Optimization attempts (problem solving)
How many times have you tried the ELISA?



Have you run a "No Primary" control?


Yes No
Do you obtain the same results every time?
Yes No

What steps have you altered?


Additional Notes:



Data:
We would appreciate if you are able to provide a copy of the data, particularly from the standard curve. This will help us to assess the results.

Read More

Answer

Further to my previous response, serum samples and other biological fluids are known to show an increase in background due to the many components found in these types of samples.

Do you have the readings for the standard and test samples as well as your detailed protocol you can send to us which we could perhaps review?

Thank you for your co-operation in this matter!

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Answer

Thank you for your reply.

I can confirm that one vial of ab83137 Anti-IGF1 antibody (Biotin)contains 50 ug. This information is provided at the top of the online datasheet.

This particular antibodyis provided lyophilised. The datasheet provides reconstitution instructions:

Centrifuge vial prior to opening. Reconstitute in sterile PBS containing 0.1% BSA to a concentration of 1.0 mg/ml.

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

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Answer

Thank you for your reply.

Reviewing the details of the experiment you are planning, I can suggest to consider which antibodies you use. As you are using a secondary antibody in this case, the secondary antibody will pick up the captureantibody on the plate and the detection antibody, because they are both from the same species. This may give high background.

If you need to use a conjugated secondary, I can suggest to use capture and detection antibodies from different species. Or to do an indirect ELISA instead.

Alternatively, we do have some conjugation kits, including forHRP. You could tryconjugating the ab9572 directly. I can suggest to read the online datasheets carefully for further information, and ensure the kit will be suitable:

ab102891EasyLink HRP Conjugation Kit (3 x 100µg HRP)
https://www.abcam.com/index.html?datasheet=102891 (or use the following: https://www.abcam.com/index.html?datasheet=102891).

ab102892 EasyLink HRP Conjugation Kit (3 x 10µg HRP)
https://www.abcam.com/index.html?datasheet=102892 (or use the following: https://www.abcam.com/index.html?datasheet=102892).

ab102889EasyLink HRP Conjugation Kit (1 x 1mg HRP)
https://www.abcam.com/index.html?datasheet=102889 (or use the following: https://www.abcam.com/index.html?datasheet=102889).

ab102890 EasyLink HRP Conjugation Kit (1 x 100µg HRP)
https://www.abcam.com/index.html?datasheet=102890 (or use the following: https://www.abcam.com/index.html?datasheet=102890).

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

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Answer

Thank you once again for your enquiries.

To follow up from my previous email, I now have some further information regarding ab83137 Anti-IGF1 antibody (Biotin)which can be used in sandwich ELISA with ab9572.

ab83137 is the same antibody as ab9572 but with the addition of the biotinylation. Therefore, the immunogen used to raise this antibody is the same IGF-I. Although there is a high level of sequence homology,we would be unable to guarantee if this antibody will detect the customersIGF-I peptide. Our antibody has a high affinity towards the natural and/or recombinant versions of the corresponding protein.

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

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