Overview

  • Product name
    Anti-Insulin antibody [3A6]
    See all Insulin primary antibodies
  • Description
    Mouse monoclonal [3A6] to Insulin
  • Host species
    Mouse
  • Specificity
    Kass for this antibody is 8.0 x 108M-1. It cross-reacts with human proinsulin. No cross-reaction with free C-peptide.
  • Tested applications
    Suitable for: Sandwich ELISA, IHC-P, ELISAmore details
  • Species reactivity
    Reacts with: Human, Pig
    Predicted to work with: Cow
  • Immunogen

    Full length native protein (purified).

  • Positive control
    • IHC-P (FFPE): Human Pancreas (Normal) tissue.
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab133281 as a replacement.

Properties

Applications

Our Abpromise guarantee covers the use of ab1965 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. Can be paired for Sandwich ELISA with Mouse monoclonal [8E2] to Insulin (HRP) (ab24455). Can be used as Capture antibody with recommended pair.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ELISA Use at an assay dependent concentration.

Target

  • 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
  • Database links
  • Alternative names
    • IDDM antibody
    • IDDM1 antibody
    • IDDM2 antibody
    • ILPR antibody
    • ins antibody
    • INS_HUMAN antibody
    • Insulin A chain antibody
    • Insulin B chain antibody
    • IRDN antibody
    • MODY10 antibody
    • Preproinsulin antibody
    • Proinsulin antibody
    • Proinsulin precursor antibody
    see all

Images

  • IHC image of Insulin staining in Human Pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1965, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

References

ab1965 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Answer

Thank you for your reply.
Please keep in mind that mouse has not been tried and we do not know if and how well the antibodies will work with this species.
I'll pass the lot numbers to the lab.

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 contacting us.

In our https://www.abcam.com/index.html?pageconfig=resource&rid=11422 we recommend carbonate/bicarbonate buffer at pH 9.6. However, the capture antibody can also be diluted with PBS, pH 7.4, and the HRP-conjugated detection MAb with PBS, 0.05% Tween-20.

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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Answer

We have used following very general protocol with this antibody pair:

1. Coating MAb, 10 μg/ml in PBS, pH 7.4, 50 μl/per well, 2h, +37 C
2. Washing with PBS, 0.05% Tween-20, 4 times
3. Inhibition with PBS, 0.05% Tween-20, 0.1% BSA (RIA grade, without insulin), 150 μl/well, 1h, +37 C
4. Washing with PBS, 0.05% Tween-20, 4 times
5. Add antigen, conc. 0.1 – 3.0 ng/ml for human insulin, 50 μl/well, in PBS, 0.05% Tween-20, 0.1% BSA; overnight, +4 C
6. Washing with PBS, 0.05% Tween-20, 6 times
7. Add HRP-conjugated detection MAb (1:800-1000 dilution), 50 μl/well, 2h, +37 C
8. Washing with PBS, 0.05% Tween-20, 6 times
9. Bound enzyme activity evaluation – TMB- H2O2 system.

Please note that this is a general protocol for insulin sandwich immunoassay. Optimal antibody and antigen working concentrations as well as incubation times should be determined separately.

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Application
ELISA
Sample
Mouse Purified protein (Insulin)
Specification
Insulin
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Type
Sandwich (Capture)

Ms. Kira Slepchenko

Verified customer

Submitted Oct 03 2012

Answer

Thank you for your phone call.
I have contacted the lab for more information about antibody dilutions and for a detailed protocol, and will let you know when I hear back.
Below, please find the discount codes for trying these antibodies with mouse samples.

DISCOUNT CODE: xxx for ab1965
DISCOUNT CODE: xxx for ab24455
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab1965 and ab24455 in mouse. Each code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

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Question
Answer

For ab24455 DISCOUNT CODE: ***

For ab1965 DISCOUNT CODE: ***

Expiration date: December 30th, 2012

I am very pleased to hear you would like to accept our offer and test ab1965 and ab24455 for useon mouse samples. Each code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for using each antibody inmouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for yoiru enquiry.

I can confirm that these two antibodies have been tested together in Sandwich ELISA system. The storage instructions can be found on the on-line product datasheets. We normally suggest our customers using the products within 6 months (Abpromise) to make sure that we can offer our full guarantee even though after proper aliquotting they can be active for months or for several years depending on the storage temperature.

Standard ELISA, sELISA can be carried out on special ELISA plates (96-well) but the size of the plates really depends on the customer's need. Thermo Scientific, Nunc, BD Biosciences sell different variety of ELISA plates.

If you need any further assistance in the future, please do not hesitate to contact me.

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Answer

Thank you for your response.

You have raised an excellent point that I will bring to our datasheet team. While we do list sandwich ELISA as a different application perhaps we could list the specific ELISA type as we do for IHC. In most cases it would read Indirect ELISA and Direct ELISA. I would like to thank you for this suggestion.

The principles behind the sandwich and indirect ELISAs (which are the application type we guarantee) are very much the same however an sandwich ELISA uses two antibodies against a specific target protein, one as capture on as detection, whereas a normal Indirect ELISA use an antibody against the species as capture and against the target protein as detection. The problems with sandwich are that the two antibodies against the same target protein may interfere with one another leading to poor or no signal. Because of this we only recommend using products that have been tested in sandwich ELISA or are matched pairs.

This information may be found on our website by searching for the protein of interest in the search bar while using the 'primary antibodies' search in the drop down menu. After the results are displayed, further narrow down the field to only those tested in sandwich ELISA by using the filter on the left hand side. In this case I was able to find 2 anti-insulin antibodies tested in sELISA (ab1965 and ab24455) as well as a few for proinsulin.

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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Answer

Thank you for contacting Abcam.

Please find the answers to your questions below:

1 - You are correct that ab1965 is the capture antibody and that ab2455 is the detection antibody. The phrase "Can be used as Capture antibody with recommended pair", simply means that you could use ab1965 as the capture antibody in a verified pair that has been shown to be successful in a sandwich ELISA. Currently however the only antibody it has been matched with is ab24455.

2 - We do have many other antibodies that may work for a sandwich ELISA, but this is the only pair that we guarantee to work.

3 - We currently do not have an non-hrp conjugated form of ab24455, but if you are interested in getting a non-conjugated form, I can check with the lab to see if it’s possible.

I hope that this information is useful and please let me know if there is anything else I can help you with.

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Answer

Good morning to you to, I hope you are having a great day. Thank you for contacting Abcam about ab1965 and ab24455. Neither of these antibodies have been tested on rat tissue and so we cannot guarantee that they will work in your application. I did an alignment between the human and rat sequences to see if they shared a high level of similarity and they are about 79% similar, so there is a chance the antibodies will recognize rat tissue. Since we cannot guarantee that these antibodies will work on rat tissue, but there is a chance it may work, we can offer you a testing discount, the details of which are below. 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 ab1965/24455 in rat tissue I will then send a discount code. This code must be issued before purchasing ab1965/24455 so please wait for my reply before ordering. 2. Purchase ab1965/24455 either by phone, fax, or online (www.abcam.com). 3. Test it in rat tissue. 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 primary antibody 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 ab1965/24455 turns out to be unsuitable for rat tissue, 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.c

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1-10 of 12 Abreviews or Q&A

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