Overview

  • Product name
    Anti-C Peptide antibody (Biotin)
    See all C Peptide primary antibodies
  • Description
    Rabbit polyclonal to C Peptide (Biotin)
  • Host species
    Rabbit
  • Conjugation
    Biotin
  • Tested applications
    Suitable for: RIA, ELISAmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Chimpanzee, Gorilla
  • Immunogen

    Synthetic peptide: EAEDLQVGQV ELGGGPGAGS LQPLALEGSL Q conjugated to KLH, corresponding to internal sequence amino acids 57-87 of Human C Peptide. Conjugated to KLH via carboxyl group

  • General notes


    C Peptide is part of the molecule of Proinsulin, that consists of three parts: C Peptide and two long strands of amino acids (called the alpha and beta chains) that later become linked together to form the insulin molecule. From every molecule of proinsulin, one molecule of insulin plus one molecule of C Peptide are produced. C peptide is released into the blood stream in equal amounts to insulin. A test of C peptide levels will show how much insulin the body is making. 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.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Thimerosal (merthiolate)
    Constituents: 50% Glycerol, PBS, pH 7.5
  • Concentration information loading...
  • Purity
    Protein G purified
  • Primary antibody notes
    C Peptide is part of the molecule of Proinsulin, that consists of three parts: C Peptide and two long strands of amino acids (called the alpha and beta chains) that later become linked together to form the insulin molecule. From every molecule of proinsulin, one molecule of insulin plus one molecule of C Peptide are produced. C peptide is released into the blood stream in equal amounts to insulin. A test of C peptide levels will show how much insulin the body is making. 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.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab48303 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
RIA Use at an assay dependent dilution.
ELISA Use at an assay dependent dilution.

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 2 antibody
    • IDDM2 antibody
    • ILPR antibody
    • ins antibody
    • INS_HUMAN antibody
    • Insulin A chain antibody
    • IRDN antibody
    • MODY10 antibody
    • Proinsulin antibody
    see all

References

ab48303 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1205485 (2 x ab20903).
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

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Answer

I agree the antibodies should detect the proteins in serum; as this is not the case so I am happy to replace these products. Would you like try new vials of these antibodies or a different antibody against the same target?
My suggestion would be trying one antibody first and if you get satisfactory results then I will send you the second.
Though ab93903 has not been tested in ELISA with ab8297 and ab9298 however lab has confirmed using this in titre test. The following procedure they used; maybe this information will help!
1 µg/ml with carbonat-buffer, NaHCO3, pH 9.6, for 2 hours by room-temperature,

100µl each well for coating.
Could you confirm how you would like to proceed?

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Question

Thanks for replying so promptly. See below for answers to your questions:

Q. Could you provide the source of protein?. Is it possible if you could provide sequence of C-peptide used?
A. We are using abcam's C-peptide calibrator (ab93903). The sequence is EAEDLQVGQV ELGGGPGAGS LQPLALEGSL Q, corresponding to amino acids 57-87 of Human C Peptide
The detection antibody recognizes the same sequence only it's conjugated to Biotin. The immunogen for the capture Ab (ab8297) is the recombinant full length protein.

Q. What was the secondary antibody used? Does this antibody worked before?
We are using an anti-mouse antibody from meso scale discovery. It's a goat polyclonal antibody that binds to heavy and light chains of mouse IgG. It's specifically designed for use in ELISA's and works very well in ELISAs as a secondary detection reagent

Q. Have you used ELISA before? Do you think ELISA reader is OK?
A. I've developed and validated lots of ELISA's from scratch and have also successfully developed and validated two multiplex ELISAs.
The plate reader is working fine as we are running other ELISA's at the moment too without problems.

Q. Was there no reading at all with serum and c-peptide?
A. We tested the detection only with the antigen (C-peptide and serum) immobilised onto the plate
The results for our diluent with skimmed milk as the blocker as are follows: The signal we got for the highest point on the standard curve was 63, the blank was 53 and for serum sample a: 55 and serum sample b: 53
The results with PBS with BSA as the blocker: The signal we got for the highest point on the standard curve was 62, the blank was 122 and for serum sample a: 67 and serum sample b: 61

When we tested the Capture Ab with the anti-species antibody we got similar results too:
For example, the signal we got for the highest point on the standard curve was 83, the blank was 50 and for serum sample a: 47 and serum sample b: 61

Many thanks,

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Answer

Thank you very much for confirming details.
The protein ab93903 has not been tested in ELISA and in combination with ab8297, ab8298 and ab48303 so we unfortunately do not hold any ELISA data for this product. This product was in fact used as blocking peptide with antibody ab14181. I have also contacted our laboratory regarding this and now waiting a reply from them. I will share their experience with you soon.
The further question I would like to ask is; were the concentration/ dilution of peptide, optimized in combination with antibodies?
Thank you very much for your cooperation.

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Question

I'm emailing to require a refund for two C-peptide antibodies we purchased (ab48303 and ab8297) which aren't working in an ELISA. We have now tried them in a direct, indirect and sandwich ELISA types. Details are as follows:

Purchase Order No: CL14137 anti-c-peptide (Biotin), ab48303
Purchase Order No: CL13977 anti c-peptide ab [1H8], ab8297

Both antibodies have been tested to work in a ELISA.

Our initial aim was to develop a sandwich ELISA even though this pair hasn't been tested for use in a sELISA. We were not able to generate a standard curve. The signal that resulted was baseline.

To determine if the problem was ab48303 we tried a direct ELISA by immobilising the purified antigen overnight (both purified C-peptide and human serum samples as positive controls were used (run both neat and diluted). Only a very low baseline signal resulted in all wells


We also tried using ab8297 in an indirect ELISA using an anti species detection Ab we know works in an ELISA. No standard curve resulted either.



For all ELISA types, we tested

1. varying concentrations of antibodies in case we weren't using enough,

2. different diluents we have successfully used for many other ELISAs with either BSA or non-fat skimmed milk for blocking

3. different incubation times

4. Human serum samples were run as positive controls in case the problem was the calibrator.



For every combination tried in each ELISA tested, the signal that resulted was baseline.





What's the next step in order to get a refund? Do we need to send you back the antibodies? We have aliquoted both of them into smaller volumes to avoid freeze/thaw issues but can re-pool if necessary.



Many thanks,

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Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody
.
I have read the details you have kindly provided and have following further questions for better understanding of the problem;
- Could you provide the source of protein?. Is it possible if you could provide sequence of C-peptide used?
- What was the secondary antibody used? Does this antibody worked before?
- Have you used ELISA before? Do you think ELISA reader is OK?
- Was there no reading at all with serum and c-peptide?
Thank you very much for your cooperation. I will be looking forward to hearing from you soon.

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Answer

Thank you very much for your interest in ab48303.
I can suggest to match this antibody with ab8297 or ab8298. Both of them are tested to work in ELISA. But it has not been tested if they work with ab48303.
To our knowledge, this antibody pair has not been tested in sandwich ELISA (sELISA).
Therefore, I can offer a discount off a future purchase if you buy a pair of antibodies now, test it in sELISA and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of the two antibodies off a future order.
If you are interested in this offer, please follow these steps:
1. Reply to this e-mail to let me know with which antibodies you would like to proceed and test them in sELISA. I will then send a discount code. This code must be issued before purchasing the antibodies so please wait for my reply before ordering.
2. Purchase the antibody pair either by phone, fax, or online (www.abcam.com).
3. Test it in sELISA.
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 product ordered and the discount code is valid for 4 months after issue. Please remember that submission of the Abreview is sufficient for the discount code to become active.
We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the antibody pair turns out to be unsuitable for sELISA, 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.com/collaborationdiscount.

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

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