Overview

  • Product name
  • Description
    Rabbit polyclonal to IGF1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, Neutralising, IHC-FoFr, ICC/IF, Sandwich ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Highly pure (>98%) recombinant hIGF-1 (human Insulin Like Growth Factor-1).

  • Positive control
    • Recombinant human IGF1 protein (ab9573) can be used as a positive control in WB.

Properties

Applications

Our Abpromise guarantee covers the use of ab9572 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
IHC-P Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. To detect hIGF-1 by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant hIGF-1 is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions.
Neutralising Use at an assay dependent concentration. To yield one-half maximal inhibition of the biological activity of hIGF-1 (5.0 ng/ml), a concentration of 0.67 - 1.0 ug/ml of this antibody is required.
IHC-FoFr Use at an assay dependent concentration. PubMed: 28754163
ICC/IF 1/200.
Sandwich ELISA Use a concentration of 0.5 - 2 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to IGF1 (Biotin) (ab83137). Allows for the detection of at least 0.2 - 0.4 ng/well of recombinant IGF1.

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

Images

  • Ab9572 staining Human normal liver parenchyma. Staining is localised to the cytoplasm.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ab9572 staining IGF1 in mouse heart tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% formalin and blocked with 10% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/1000 in 10% goat serum) for 24 hours at 4°C. An Alexa Fluor® 594-conjugated goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-IGF1 antibody (ab9572) at 1/2000 dilution

    All lanes : Human vascular smooth muscle whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Donkey Anti-Rabbit IgG - HRP at 1/10000 dilution

    Developed using the ECL technique.

    See Abreview

  • ICC/IF image of ab9572 stained human embryonic stem cells. The cells were fixed in methanol and then incubated in 10% BSA for 1 hour to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9572, 1/200) overnight at +4°C. The secondary antibody (red) was an Alexa Fluor® 568 goat anti-rabbit used at a 1/400 dilution.

    See Abreview

References

This product has been referenced in:
  • Ma M  et al. Preeclampsia is associated with hypermethylation of IGF-1 promoter mediated by DNMT1. Am J Transl Res 10:16-39 (2018). IHC-P ; Human . Read more (PubMed: 29422991) »
  • Hsu PY  et al. MicroRNA let-7g inhibits angiotensin II-induced endothelial senescence via the LOX-1-independent mechanism. Int J Mol Med 41:2243-2251 (2018). Read more (PubMed: 29393358) »
See all 38 Publications for this product

Customer reviews and Q&As

1-10 of 26 Abreviews or Q&A

Application
Flow Cytometry
Sample
Mouse Cell (EL4 T cells tumor line and primary hepatocytes)
Permeabilization
No
Gating Strategy
Positive cells for FITC
Specification
EL4 T cells tumor line and primary hepatocytes
Preparation
Cell harvesting/tissue preparation method: Mice were dissected to obtain the liver. The liver was then mashed to obtain hepatocytes.
Sample buffer: PBS with 10% bovine serum
Fixation
none

Safa Mohamad

Verified customer

Submitted May 12 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse neuron)
Permeabilization
Yes - triton X-100
Specification
mouse neuron
Blocking step
BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 28 2015

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
None
Specification
Brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 15 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step
Heat mediated
Sample
Mouse Tissue sections (Heart)
Specification
Heart
Permeabilization
No
Fixative
10% Formalin

Abcam user community

Verified customer

Submitted Jul 15 2014

Question
Answer

To our knowledge, this IGF antibody has not been tested in flow cytometry and our guarantee does not cover this application of the antibody. However it has been tested and shown to work in a similar application, immunocytochemistry, after methanol fixation of the cells. For flow cytometry, your cells will need to be fixed and permeabilized, as IGF is completely intracellular until it is secreted.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cultured Cells (human embryonic stem cells)
Specification
human embryonic stem cells
Fixative
Methanol
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted May 13 2013

Answer

Thank you for contacting us.

The lab sent me the following information:

The immunizing antigen was produced in E.coli.
The purification of this antibody is done using an antigen affinity column.
This is all the information we have available.

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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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.

Read More

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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Read More
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

1-10 of 26 Abreviews or Q&A

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