Overview

  • Product name
    Anti-GLP1 (amidated) antibody [8G9]
  • Description
    Mouse monoclonal [8G9] to GLP1 (amidated)
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-P, IHC-Fr, Sandwich ELISA, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Guinea pig, Cow, Human, Pig, Common marmoset
    Predicted to work with: a wide range of other species
  • Immunogen

    Synthetic amide peptide identical to human, bovine, guinea pig, mouse, rat and all other examined mammalian species: H-

    HAEGTFTSNVSSYLEGQAAKEFIAWLVKGR

    -NH2, corresponding to amino acids 7-36 of GLP 1

  • Epitope
    C-terminal epitope of GLP-1(7-36)amide
  • Positive control
    • IHC-P/IHC-Fr: rat colon and rat pancreas

Applications

Our Abpromise guarantee covers the use of ab26278 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 1/2000. Fix in 4% paraformaldehyde in 0.1 phosphate buffer, pH 7.4 overnight at 4°C.
IHC-Fr 1/2000. Best performance has been seen in frozen sections.
Sandwich ELISA Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Mouse monoclonal [4F3] to GLP1 (ab23472).

We recommend that ab26278 is used as capture antibody. ab26278 binds to the c-terminus and presents the mid-molecular epitope to the ab23472 detection antibody, so it has easy access to it. For the same reason we do not recommend swapping the pair, as the binding of the mid molecular epitope to the coat will not present the c-terminus epitope to the ab26278 antibody in a very favourable manner.

ELISA Use at an assay dependent concentration.

Target

  • Function
    Glucagon plays a key role in glucose metabolism and homeostasis. Regulates blood glucose by increasing gluconeogenesis and decreasing glycolysis. A counterregulatory hormone of insulin, raises plasma glucose levels in response to insulin-induced hypoglycemia. Plays an important role in initiating and maintaining hyperglycemic conditions in diabetes.
    GLP-1 is a potent stimulator of glucose-dependent insulin release. Play important roles on gastric motility and the suppression of plasma glucagon levels. May be involved in the suppression of satiety and stimulation of glucose disposal in peripheral tissues, independent of the actions of insulin. Have growth-promoting activities on intestinal epithelium. May also regulate the hypothalamic pituitary axis (HPA) via effects on LH, TSH, CRH, oxytocin, and vasopressin secretion. Increases islet mass through stimulation of islet neogenesis and pancreatic beta cell proliferation. Inhibits beta cell apoptosis.
    GLP-2 stimulates intestinal growth and up-regulates villus height in the small intestine, concomitant with increased crypt cell proliferation and decreased enterocyte apoptosis. The gastrointestinal tract, from the stomach to the colon is the principal target for GLP-2 action. Plays a key role in nutrient homeostasis, enhancing nutrient assimilation through enhanced gastrointestinal function, as well as increasing nutrient disposal. Stimulates intestinal glucose transport and decreases mucosal permeability.
    Oxyntomodulin significantly reduces food intake. Inhibits gastric emptying in humans. Suppression of gastric emptying may lead to increased gastric distension, which may contribute to satiety by causing a sensation of fullness.
    Glicentin may modulate gastric acid secretion and the gastro-pyloro-duodenal activity. May play an important role in intestinal mucosal growth in the early period of life.
  • Tissue specificity
    Glucagon is secreted in the A cells of the islets of Langerhans. GLP-1, GLP-2, oxyntomodulin and glicentin are secreted from enteroendocrine cells throughout the gastrointestinal tract. GLP-1 and GLP-2 are also secreted in selected neurons in the brain.
  • Sequence similarities
    Belongs to the glucagon family.
  • Post-translational
    modifications
    Proglucagon is post-translationally processed in a tissue-specific manner in pancreatic A cells and intestinal L cells. In pancreatic A cells, the major bioactive hormone is glucagon cleaved by PCSK2/PC2. In the intestinal L cells PCSK1/PC1 liberates GLP-1, GLP-2, glicentin and oxyntomodulin. GLP-1 is further N-terminally truncated by post-translational processing in the intestinal L cells resulting in GLP-1(7-37) GLP-1-(7-36)amide. The C-terminal amidation is neither important for the metabolism of GLP-1 nor for its effects on the endocrine pancreas.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • GCG antibody
    • GLP-1 antibody
    • GLP-1(7-36) antibody
    • GLP-1(7-37) antibody
    • GLP-2 antibody
    • GLUC_HUMAN antibody
    • Glucagon-like peptide 2 antibody
    • GRPP antibody
    • Incretin hormone antibody
    • OXM antibody
    • OXY antibody
    see all

Images

  • ab26278 (1µg/ml) staining GLP1 in Human pancreas, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

References

This product has been referenced in:
  • Zhang Y  et al. Differential Effects of Linagliptin on the Function of Human Islets Isolated from Non-diabetic and Diabetic Donors. Sci Rep 7:7964 (2017). Read more (PubMed: 28801559) »
  • Cavin JB  et al. Differences in Alimentary Glucose Absorption and Intestinal Disposal of Blood Glucose After Roux-en-Y Gastric Bypass vs Sleeve Gastrectomy. Gastroenterology 150:454-64.e9 (2016). Read more (PubMed: 26481855) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Answer


The lab confirmed that if one considers the GLP1 pre-protein to be the aa 1-37 amidated form of GLP1, then that is what the antibody recognizes.

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Question
Answer

From review of previous discussion with the originator of this product, it appears that there should be no reactivity with glucagon. I have also BLAST-ed the immunogen and aligned with glucagon. The immunogen HAEGTFTSNVSSYLEGQAAKEFIAWLVKGR, corresponding to amino acids 7-36 of GLP 1, has only a 47% sequence homology with glucagon and we would not expect this antibody to recognize this. While targeted specifically for GLP1, this antibody may react with any glucagon preproprotein present in your sample as this would contain both GLP1 and glucagon, though this would depend upon epitope presentation for GLP1.

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Answer

Thank you for contacting us.
I have had a look at the products that we have available in our catalogue and I recommend using the following validated antibody pairs for sandwich ELISA (sELISA):
1. Glucagon Like Peptide-1 (active)
I have found two different antibody pairs that have been validated for use in sELISA:
Capture: ab23472 (www.abcam.com/ab23472) and Detection: ab121080(www.abcam.com/ab121080).
or
Capture: ab26278 (www.abcam.com/ab26278) and Detection: ab121072(www.abcam.com/ab121072).
ab26278 is specific for the amidated C terminus of the peptide and does not react with GLP 1 (7-37). It reacts with the amidated C-terminus of GLP 1 (7-36) amide, GLP-1(9-36) and GLP 1 (1-36) amide.
2. Peptide YY
Capture: 121069 (www.abcam.com/ab121069) and Detection: ab121070(www.abcam.com/ab121070).
3. Human Ghrelin
Capture: 121066 (www.abcam.com/ab121066) and Detection: ab121067(www.abcam.com/ab121067).
Attached is a word document with the 4 “standard curves” from the 4 assays. Please note however that these assays have not been optimized and it is very likely that with optimization, you will be able to retrieve a better assay performance than what has been shown here.
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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Question
Answer

Thank you for contacting Abcam.


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.

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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Question
Answer

Thank you for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending a free of charge vial of this antibody on the order ***, which should arrive on Friday.

Please keep me updated about the results with this antibody, and let me know if there is anything else that we can do for you. Have a great week!

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Answer

Thank you for your patience.


Please find the original reference for this antibody attached. The product mentioned in the paper is HYB 147-6.


Unfortunately, theoriginator does not have any more data that can be disclosed. However, as HYB 147-6 (ab26278) reacts with the amidated GLP-1 – and this area of Glucagon is not amidated, there is no cross reaction.


I hope this information will be helpful.

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Answer

Thank you for your enquiry.


I have checked with the originator of the product and according totheir data,no cross reaction with glucagon was detected. This product is specific for GLP1 protein expression.


I hope this information would be useful for you.

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Answer

Thank you for contacting us.

ab26278 would work with Mouse tissue and in IHC-Fr. Please be advised that this is a primary antibody not the secondary antibody. The compatible FITC conjugated secondary antibodies are

ab97239, ab98692 and ab115588.

"Cat # 1072-1002" this is not an Abcam catalogue number so please recheck it.

For mouse on mouse staining, please follow the guidelines given inbelow link;

https://www.abcam.com/ps/pdf/protocols/mouse_on_mouse.pdf

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 your enquiry.

I am sorry I am not able to find catalog number 1072-1002 on our system. Our product numbers always start with ab. We do have the following goat anti mouse IgG FITC conjugated secondary antibody, tested in IHC-Fr, which should be suitable for use with ab26278.

ab7064Goat anti-Mouse IgG H&L (FITC) secondary antibody
FITC conjugated
Flow Cyt, IHC-P, ELISA, IM, ICC/IF, IHC-Fr
https://www.abcam.com/index.html?datasheet=7064 (or use the following: https://www.abcam.com/index.html?datasheet=7064).

I would suggest to bear in mind that the excitation and emission spectra for FITC are very similar to Alexa 488, and so I would not advise using these two conjugates together in duel staining as you will not be able to distinguish between them.

If you would prefer to use a suitable secondary antibody with a different fluorophore, then I can suggest considering the following which has been tested in IHC-Fr:

ab7058Donkey polyclonal Secondary Antibody to Mouse IgG - H&L (TRITC ), pre-adsorbed
TRITC conjugated
https://www.abcam.com/index.html?datasheet=7058 (or use the following: https://www.abcam.com/index.html?datasheet=7058).

I can recommend to review the flourophores you wish to use, and if you require any further help please let me know.

This particular secondary (ab7058) is also cross absorbed, which means it is less likely to cross react. However, as you mention, there is always the possibility of crossreactivity when an anti-mouse secondary is used on mouse tissue. I can recommend to review the following page from our website which provides some useful tips:

https://www.abcam.com/index.html?pageconfig=resource&rid=11465

I hope this will be helpful to you. If you have any further questions, please do not hesitate tocontact us.

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Answer

Thank you for contacting Abcam.

The peptide used when this antibody was developed was made for the immunization, therefore we do not have it commercially available.

I am sorry that I could not be more help in this matter.

If there is anything else I can help you with, please let me know.

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1-10 of 13 Abreviews or Q&A

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