Overview

  • Product name
    Anti-Insulin + Proinsulin antibody [D3E7] (Biotin)
    See all Insulin + Proinsulin primary antibodies
  • Description
    Mouse monoclonal [D3E7] to Insulin + Proinsulin (Biotin)
  • Host species
    Mouse
  • Conjugation
    Biotin
  • Tested applications
    Suitable for: IHC-Fr, Sandwich ELISA, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Pig
  • Immunogen

    Full length protein (Human).

  • General notes
    For concentration (OD280nm, E0.1% = 1.4). Affinity constant Kd = 6.3 x 10E-8M.

Properties

Applications

Our Abpromise guarantee covers the use of ab20756 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-Fr Use at an assay dependent concentration.
Sandwich ELISA Use at an assay dependent concentration.

Can be paired for sandwich ELISA with Mouse monoclonal [D6C4] to Insulin + Proinsulin (ab8304).

ELISA Use at an assay dependent concentration. Can be paired for ELISA with Mouse monoclonal [D6C4] to Insulin + Proinsulin (ab8304).

Target

Images

  • ab20756 staining Insulin + Proinsulin in human pancreas tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100 and blocked for 1 hour at 25°C. Samples were incubated with primary antibody (1/100) for 1 hour at 25°C followed by incubation with Cy3®-conjugated streptavidin for 30 minutes in the dark at room temperature. Nuclei were counterstained with DAPI.

    See Abreview

References

This product has been referenced in:
  • Kuhn C  et al. Human CD3 transgenic mice: preclinical testing of antibodies promoting immune tolerance. Sci Transl Med 3:68ra10 (2011). IHC-Fr ; Mouse . Read more (PubMed: 21289272) »
  • Yang RY  et al. Ablation of a galectin preferentially expressed in adipocytes increases lipolysis, reduces adiposity, and improves insulin sensitivity in mice. Proc Natl Acad Sci U S A 108:18696-701 (2011). ELISA ; Mouse . Read more (PubMed: 21969596) »
See all 2 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Answer

This antibody is already conjugated to biotin and therefore does not need a secondary antibody for detection.
What you will need is avidin or streptavidin conjugated to an enzyme or fluorochrome that will bind to multiple sites on the biotinylated primary antibody. We have several streptavidin proteins in our catalogue conjugated to an enzyme or fluorochrome. The conjugate you choose will depend on your detection system.
For more information, please visit:
https://www.abcam.com/index.html?pageconfig=resource&rid=12922
https://www.abcam.com/index.html?pageconfig=resource&rid=12723
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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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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Answer

Thank you for your inquiry. 
The antibody pair that we use in the Human Insulin ELISA Kit (ab100578) is capture antibody (ab8304) and detection antibody (ab20756). 
https://www.abcam.com/insulin--proinsulin-antibody-d6c4-ab8304.html
https://www.abcam.com/insulin-proinsulin-antibody-d3e7-biotin-ab20756.html
We have not specifically tested human analogs with this kit but based on sequence similarity, we would expect the kit to recognize these as well.  However, we wouldn't guarantee it since it hasn't been specifically tested.
I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for your further enquiry.

I am sorry to confirm that as far as we are aware, ab8304 and ab20756 insulin antibodies have never been tested in ELISPOT. All tested and guaranteed applications are specified on our datasheets, and these are updated as soon as any new information is brought to our attention.

If you would like to test these antibodies in ELISPOT, please contactmeprior to the purchase by replying to this message as you may be eligible for our testing discount program.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact me for any further advice or information in this regard.

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Answer

Thank you for your enquiry which has been forwarded to the scientific support team. We can be contacted at mailto:technical@abcam.com, or directly from the website.

I can confirm that ab8304 can be paired for Sandwich ELISA with Mouse monoclonal [D3E7] to Insulin + Proinsulin (Biotin) (ab20756).

In this case, it is ok to use a mouse monoclonal for the coating antibody and detection antibody, because the detection antibody is directly conjugated to biotin.

(Without the biotin, if you needed to use a conjugated secondary anti mouse antibody, then this would not work as the secondary would also detect the coating antibody, which is covering the whole plate).

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

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Answer

Thank you for contacting us. Yes, the clones D6C4 and D3E7 have been validated for use as a sandwich ELISA pair and will be guaranteed for that purpose. Because the detection antibody you are choosing is already biotinylated, it will not be an issue that the two antibodies are from the same host animal. This would only be a problem if you were going to detect the signal with an anti-mouse secondary antibody, because it would react with both the capture and detection antibodies. I hope this helps, please let me know if you need need any additional information.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (pancreas)
Specification
pancreas
Fixative
Acetone
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 15 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Pancreas)
Specification
Pancreas
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.25% Titox x100
Blocking step
Rouche Blocking reagent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%

Abcam user community

Verified customer

Submitted Nov 03 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Pancreas)
Permeabilization
Yes - 0.25% Triton x100
Specification
Pancreas
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 21 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (pancreas)
Specification
pancreas
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3%

Dr. Anahi Chavarria

Verified customer

Submitted Sep 15 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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