Overview

  • Product name
  • Description
    Rabbit polyclonal to CGRP
  • Host species
    Rabbit
  • Specificity
    The immunogen used to raise this antibody has 78% homology with the human CGRP protein. Previous batches of ab47027 have shown to work in human tissue. Please contact Abcam Scientific Support for more information.
  • Tested applications
    Suitable for: IHC-Pmore details
  • Species reactivity
    Reacts with: Rat
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Rat CGRP.

    Read Abcam's proprietary immunogen policy (Peptide available as ab47101.)

  • Positive control
    • IHC-P: FFPE rat dorsal root ganglion, spinal cord and rat thyroid tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab47027 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 Use a concentration of 0.1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Relevance
    CGRP is present in C-cells of the thyroid and in central and peripheral nerves. CGRP has several important physiologic roles: it is a potent vasodilator, and can affect the force and rate of heart beat; it can modulate acetylcholine receptor function at the neuromuscular junction; it has been demonstrated to block tolerance to morphine; and it can modulate antigen presentation in Langerhans cells in the skin. Despite these important physiologic functions, therapeutic strategies using CGRP have been impeded due to the lack of a cloned CGRP receptor with which ligands could be developed.
  • Cellular localization
    Endoplasmic reticulum and Secreted
  • Database links
  • Alternative names
    • Calcitonin/calcitonin related polypeptide alpha antibody
    • Alpha type CGRP antibody
    • Beta type CGRP antibody
    • CALC1 antibody
    • CALC2 antibody
    • CALCA antibody
    • CALCB antibody
    • Calcitonin 1 antibody
    • Calcitonin 2 antibody
    • Calcitonin gene related peptide I antibody
    • Calcitonin gene related peptide II antibody
    • Calcitonin related polypeptide alpha antibody
    • Calcitonin related polypeptide beta antibody
    • CGRP antibody
    • CGRP I antibody
    • CGRP II antibody
    • CGRP1 antibody
    • CGRP2 antibody
    • CT antibody
    • Katacalcin antibody
    • KC antibody
    see all

Images

  • IHC image of CGRP staining in a Rat normal thyroid 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 ab47027, 0.1µg/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.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • IHC image of CGRP staining in rat dorsal root ganglion 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 ab47027, 5µg/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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • IHC-P image of CGRP staining on Rat spinal cord sections using ab47027 (1:400). The sections were deparafinized and subjected to heat mediated antigen retrieval. The sections were then blocked using 1% BSA at 21°C for 10 min. The primary antibody was used 21°C for 2 hours.

    See Abreview

References

This product has been referenced in:
  • Liu M  et al. Amylin and diabetic cardiomyopathy - amylin-induced sarcolemmal Ca2+ leak is independent of diabetic remodeling of myocardium. Biochim Biophys Acta 1864:1923-1930 (2018). ICC/IF . Read more (PubMed: 29066284) »
  • Lian YL  et al. Elevated expression of transient receptor potential vanilloid type 1 in dorsal root ganglia of rats with endometriosis. Mol Med Rep 16:1920-1926 (2017). Read more (PubMed: 28627595) »
See all 10 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain tissue)
Specification
Brain tissue

Abcam user community

Verified customer

Submitted Apr 07 2017

Answer

Thank you for contacting us. I would be very happy to refund these products for you. I have only found orders for 1 vial of this product associated with your user account. This is for order number xxxxx. Could you please provide the Abcam order confirmation number/s for any additional vials of ab47027 which you have received? I look forward to your reply. 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

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
IHC - Wholemount
Sample
Mouse Tissue (tongue)
Specification
tongue

Abcam user community

Verified customer

Submitted Sep 25 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Spinal cord)
Specification
Spinal cord
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Mr. Carl Hobbs

Verified customer

Submitted Sep 06 2012

Answer



I am very pleased to hear your customer would like to test ab47027 Anti-CGRP antibody in rabbit. This code will give them1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for rabbit and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. 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: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for contacting us.


I performed a protein alignment with the target immunogens and the rabbit protein sequences for the antibodies. We recommend that alignment should be over 85% to predict that an antibody will detect in a different species.

Here are the results, similarities are in %:

ab47027 Anti-CGRP antibody: 85%

ab14184 Anti-Substance P antibody [SP-DE4-21]: 100%

ab106291 Anti-Substance P antibody: 100%

ab2318: As mentioned in the Scientific Support section, PGE2 is a lipid rather than a protein and is therefore not species specific. There should not be any particular problems using this assay to detect PGE2 from rabbit. Optimization for different sample types will have to be done by the end user. Please keep in mind that since PGE2 is a lipid, extraction buffers containing detergents (for preparing tissue samples) are not recommended. Mechanical disruption in a buffer such as PBS is recommended instead. For serum and plasma, simple dilution should be sufficient.


I can offer a discount off a future purchase if your customer buys one or more of the antibodiesnow, test it in rabbit 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.


If they are interested in this offer, please let me know.

Read More

Answer

Thank you for your enquiry.

I can confirm that the immunogen for ab47027spans part of the signal peptide and the first propeptide so it should only detect full length unprocessed CGRP.

The immunogen sequence is:
##############



The WB images we obtain when using this antibody generate a ˜15 kDa band which would also suggest it's the full length unprocessed protein that is being detected. I am sorry we have not tested it specifically for detection of the shorter mature form of CGRP in the laboratory, so we are not able to provide data to actually confirm that it would not detect.

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

Read More

Answer

Thank you for taking the timetotry the suggestions.

I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody again. I appreciate the time spent in the laboratory and understand the concerns. It is regrettable the results have not been successful. The quality of our products is important to us and I would like to reassure you that we willinvestigate this case further.

Of course, I would be pleased to arrange for a replacement with another catalog number. We have a number of anti-CGRP antibodies available (such as ab91007) of which the customer could choose from. Otherwise, I can offer any other primary antibody of the customer's choice, or a credit note in compensation.

I apologise for the inconvenience and thank you for your cooperation. I look forward to hearing from you with details of how your customerwould like to proceed.

Read More

Answer

Thank you and your customer for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody. The details your customer has kindly provided will enable us to investigate this case for your customer and this is also helpful in our records for monitoring of quality. Reviewing this case, I would like to offer some suggestions to help optimise the results from ab47027: 1.) To reduce un-specific staining I suggest to increase the blocking time to 1 hour. 2.) For the most specific staining and also a full saturation of the tissue with the primary antibody, I strongly recommend that the incubation time is increased to over night at 4C. 3.) To determine that the background staining is not due to the secondary antibody, I suggest to run a no primary control. 4.) I also recommend to make sure that the samples does not dry out since these samples than produce high background. I would also appreciate if your customer can confirm some further details: 1.) Unfortunately, I received the wrong image (WB and different antibody). Could you please provide an image of the IHC-P experiment? 2.) What sample/ tissue was used for this experiment? 3.) It is stated that the experiment was repeated three times- which antigen retrieval methods were used and what were the timings of them? 4.) For how long was the sample fixed in 10% Formalin? We are happy to offer this technical support. In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. I hope this information is helpful, thank you and your customer for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Question
Answer

Thank you for your patience while I looked into the lysate used for testing this antibody.  Here is the lab's reply: "The lysate we use for testing this antibody is actually supplied by a collaborator.  I don’t have a lot of information about the lysate but it is supplied to us in RIPA buffer so the customer might like to try an extraction using that if he hasn’t already. We dilute the lysate stock further in RIPA buffer prior to use and also add DTT and loading buffer." I hope this information helps.  Please do not hesitate to contact us if you need anything further!

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1-10 of 15 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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