Overview

  • Product name
    Anti-Insulin + Proinsulin antibody [D6C4]
    See all Insulin + Proinsulin primary antibodies
  • Description
    Mouse monoclonal [D6C4] to Insulin + Proinsulin
  • Host species
    Mouse
  • Specificity
    This antibody is specific for insulin and proinsulin.
  • Tested applications
    Suitable for: Electron Microscopy, IP, ELISA, IHC-Fr, ICC/IF, Sandwich ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Pig
  • Immunogen

    Human insulin.

  • General notes

    Kd for this antibody is 8.1 x 10-8M.

    Detection of both insulin and proinsulin.

Properties

Applications

Our Abpromise guarantee covers the use of ab8304 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
Electron Microscopy Use at an assay dependent concentration. PubMed: 19628478
IP 1/100.
ELISA Use a concentration of 10 µg/ml.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 24614166
Sandwich ELISA Use a concentration of 10 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [D3E7] to Insulin + Proinsulin (Biotin) (ab20756) and Mouse monoclonal [D3E7] to Insulin + Proinsulin (HRP) (ab28063). Use at a concentration of 10 µg/ml, 50µL per well. Can be used as Capture antibody with recommended pair(s).
IHC-P 1/1250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

Images

  • ab8304 at 1/1250 staining developing mouse pancreas tissue sections (E14/15) by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in Tris buffer was performed. The tissue was then blocked and incubated with the antibody for 2 hours. A biotinylated goat anti-mouse IgG antibody was used as the secondary.

    See Abreview

  • ab8304 used on human pancreas. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in Tris buffer was performed. The tissue was then blocked and incubated with the antibody for 2 hours. A biotinylated goat anti-mouse IgG antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Kim J  et al. HX-1171 attenuates pancreatic ß-cell apoptosis and hyperglycemia-mediated oxidative stress via Nrf2 activation in streptozotocin-induced diabetic model. Oncotarget 9:24260-24271 (2018). Read more (PubMed: 29849938) »
  • Yang KC  et al. Low-adhesive ethylene vinyl alcohol-based packaging to xenogeneic islet encapsulation for type 1 diabetes treatment. Biotechnol Bioeng 115:2341-2355 (2018). Read more (PubMed: 29777589) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 36 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Bos taurus Serum (Blood plasma (EDTA))
Specification
Blood plasma (EDTA)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Type
Sandwich (Detection)

Julie Kim

Verified customer

Submitted Nov 17 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (pancreatic islet cells)
Loading amount
50 µg
Specification
pancreatic islet cells
Gel Running Conditions
Non-reduced Denaturing (4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Sarah Wang

Verified customer

Submitted May 15 2013

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

I am very pleased to hear you would like to use ab8304 in WB and send us an image of your results.

This code will give you $367 off your next order before the expiration date. To redeem this offer, please submit an Abreview with the image for WB 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.

The code will be active once the Abreview with the image 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. Remember if the product does not work you will be covered by our Abpromise guarantee and eligible for a full refund or replacement.

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/abtrial.

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Question
Answer

Thank you for contacting us. I found in our records a note stating that western blotting was added to the list of tested applications for the insulin/pro-insulin antibody ab8304 based on a single report from a customer. Unfortunately, the customer did not send an image of their blot. They briefly described their protocol as follows:

Dilution: 1/2000. Use under non reducing condition. A customer reported successfully staining a nitrocellulose blot of mammalian cell culture media transferred from a non-reducing SDS-PAGE gel. The blot was blocked with 4% skim milk in PBS.

Please let me know if you have any questions.

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Answer

Thank you for your enquiry.



Please find enclosed the requested documents as attachments.



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

Read More

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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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (MIN6 cell line)
Total protein in input
600 µg
Specification
MIN6 cell line
Treatment
RIPA buffer with SDS for 5 min
Immuno-precipitation step
Protein A

Yuichi Tsuchiya

Verified customer

Submitted Jul 05 2012

Answer

Thank you for your patience. My colleague has just added the biotinylated form of this clone to our catalog as ab133958. I hope this helps, please let me know if you need any additional assistance.

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Answer

Thank you for contacting us.

While we do not currently have a biotin conjugated form of ab8304 in our catalog, I am looking into the possibility of adding one for you. I will let you know as soon as I have any information about whether this will be possible or not. We do have another Insulin + Proinsulin antibody that is already biotinylated: ab20756. Alternatively, if you would like to biotin conjugated ab8304, we do sell conjugation kits that would be suitable for that purpose, such as ab102867. I hope this helps, please let me know if you need any additional information or assistance and I will be happy to help you further.

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1-10 of 36 Abreviews or Q&A

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

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