Overview

  • Product name
    Anti-Cy5 antibody [CY5-15]
  • Description
    Mouse monoclonal [CY5-15] to Cy5
  • Host species
    Mouse
  • Tested applications
    Suitable for: Dot blot, ELISA, IP, ICC, IHC-FoFr, Flow Cytmore details
  • Immunogen

    Other Immunogen Type corresponding to Cy5. A mixture of proteins labelled with Cy5.

  • Positive control
    • IHC-Free Floating: Rat brain tissue. Flow Cytometry: Purified human CD4+ cells.
  • General notes

    Monoclonal antibodies to Cy5 are an important tool for Cy5 signal amplification in assays where biomolecules are labelled with Cy5.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purification notes
    Purified Immunoglobulin
  • Primary antibody notes
    Monoclonal antibodies to Cy5 are an important tool for Cy5 signal amplification in assays where biomolecules are labelled with Cy5.
  • Clonality
    Monoclonal
  • Clone number
    CY5-15
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab52061 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
Dot blot Use a concentration of 1 - 2 µg/ml.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-FoFr 1/50.
Flow Cyt Use at an assay dependent concentration. PubMed: 24376610

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Relevance
    Cy3 and Cy5 are reactive watersoluble fluorescent dyes of the cyanine dye family used for labelling proteins, peptides, DNA, RNA, and other biomolecules. They are usually synthesized with reactive groups on either one or both of the nitrogen side chains so that they can be chemically cross-linked to nucleic acids or protein molecules. Thay are small, pH insensitive (between pH 3 and pH 10), soluble in aqueous solution and have tolerance to DMSO. They are more photostable than fluorescein, have high molar extinction coefficients and favorable quantum yields. They are used in many different biological assays, including DNA microarrays, protein microarrays, two-dimensional protein analysis, fluorescence resonance energy transfer, and immunocytochemistry. Cy3 is excited maximally at 550 nm and emits maximally at 570 nm, in the green part of the spectrum. Cy5 is excited maximally at 649 nm and emits maximally at 670 nm, in the red part of the spectrum.

Images

  • αOrai1 internalization in purified human CD4+ cells was measured following incubation at 37°C by flow cytometry using 2 µg/mL αOrai1-AF647 and cell surface detection with biotinylated ab52061 followed by streptavidin-BV421. Samples at 4°C were analyzed in parallel as negative controls for internalization. Data is calculated as percentage of total CD4+ cells and is the average of three donors. D) Representative flow cytometry plots of surface bound αOrai1 compared to total internalized and surface αOrai1. Panel D is representative of two experiments.

  • ab52061 staining Cy5 in rat brain tissue by Immunohistochemistry (Free Floating sections). Samples were incubated with the primary antibody at a 1/50 dilution for 20 hours at 20°C. A biotin-conjugated horse anti-mouse IgG, rat-adsorbed monoclonal was used as secondary antibody at a 1/200 dilution.

    See Abreview

References

This product has been referenced in:
  • Rossiello F  et al. DNA damage response inhibition at dysfunctional telomeres by modulation of telomeric DNA damage response RNAs. Nat Commun 8:13980 (2017). Read more (PubMed: 28239143) »
  • Bolós M  et al. Soluble Tau has devastating effects on the structural plasticity of hippocampal granule neurons. Transl Psychiatry 7:1267 (2017). IHC ; Mouse . Read more (PubMed: 29217824) »
See all 3 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer



Leider sehe ich ein paar Probleme bei diesem Protokoll:

ab52061 wurde nicht getestet ob er auch Cy5.5 erkennt. Das Immunogen war eine Mischung aus Cy5 konjugierten Proteinen.

Falls ab52061 Cy5.5 erkennen würde, hätten wir ein Problem mit dem sekundären Antikörper, da ab52061 aus Maus stammt und der passende sekundäre dann auch endogene Antikörper im Gewebe erkennen würde.

Ich schlage deshalb vor einen Antikerer zu nehmen, der getestet ist für Cy5.5 und aus Kaninchen stammt benutzen. Leider haben wir keinen solchen Antiköper im Sortiment im Moment.

Vielleicht kann die Biocompare Datenbank Ihnen da weiterhelfen:

www.biocompare.com
http://www.biocompare.com/ProductCategories/2045/Antibodies.html

Wenn Sie solch einen Antikörper finden, könnte dann eines der folgenden Kits eingesetzt werden:

ab94734

https://www.abcam.com/index.html?datasheet=94734 (or use the following: https://www.abcam.com/index.html?datasheet=94734).

ab94737

https://www.abcam.com/index.html?datasheet=94737 (or use the following: https://www.abcam.com/index.html?datasheet=94737).

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Question
Answer


Unfortunately, ab52061 has not been tested against either Cy5.5 nor Cy7. Based on the dissimilarity in chemical structure of Cy5.5 and Cy7 vs Cy5, it seems unlikely that ab52061 will recognize either Cy5.5 or Cy7.

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Question

I'm a neuroscience researcher working on neural anatomical tract tracing and imaging. My work is mainly about inject fluorescent tracers into animal brain in vivo and let the tracer undergo anterograde or retrograde transport, to label neural tissues. So one of the tracer which providing me very good quality is Alexa 647 (http://products.invitrogen.com/ivgn/product/D22914), from invitrogen. What is more complicated is i also want to check the tissue under electron microscope, so I will need to further treated the tissue (normally 80um brain slices) with OsO4 (osmium tetroxide) and embed it into durcupan. Then I will cut ultra-thin section out of it, typically in the range of 60˜80nm. So ideally i want to visulize Alexa 647 signal on those treated ultra-thin sections, but at this level normally the fluorescent was to a large extend killed or diminished. With some other tracer (eg:Texas Red or Lucifer Yellow), we tried commercial available antibody to target the tracer in those ultra-thin sections and re-visulize it, and sometimes works. However it seems there's very rare primary antibody which could targeting Alexa 647, on the market.

So I went across your product ab52060 and ab52061 (they seems developed to target Cy5), and found in their description they can also target Alexa 647. It will helps me a lot if you can give me some more information about these two antibodies, like how specific it can bind to Alexa 647 (preferably if you can give me some images or reference that tried this antibody on Alexa 647)? And do you think it can somehow works after the possible transformational change of the fluorophore from my treatment?

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Answer

Thank you for contacting us.

Unfortunately we do not have any images of product ab52061 and ab52060 with Alexa 647. Alexa 647 is based on the structure of Cy5 and therefore is structurally very similar. As our data sheet states this antibody cross-reacts with Alexa 647, it is likely this was tested during R&D stages.

The only reference that we are aware of for product ab52060 is:
Myosin-Va Mediates RNA Distribution In Primary Fibroblasts From Multiple Organs. Salerno, V.P., et al. Cell Motil. Cytoskeleton 65, 422-33, (2008)

We have not tested either antibody with denatured Cy5 or Alexa 647. If the structure is changed quite drastically I think it likely that the antibodies would not recognise the molecule. If the changes to the structure are minor there may be a chance of recognition occurring. As ab52060 has wider reactivity (Cy5 and Cy3 related molecules), this might be more likely than ab52061 to be tolerant to slight changes to target molecule structure.

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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Answer

Thank you for your inquiry.

Unfortunately I am sorry to confirm we have no data or experimental information to verify whether ab52061 Cy5 antibody [CY5-15]cross reacts with Cy5.5. The Cy5 and Cy5.5 structures are quite similar and so the possibility of cross reactivity cannot be ruled out. However, we have not data to confirm or guarantee this.

I am sorry we also do not have any antibodies against Cy5.5 available in our catalog. Therefore, I would like to recommend checking the Biocompare website which has an excellent antibody search facility that includes many suppliers. The links are:

http://www.biocompare.com
http://www.biocompare.com/ProductCategories/2045/Antibodies.html

I am sorry we do not have the product you require on this occasion. However, I hope this information is nevertheless helpful to you. Please do not hesitate to contact me if you have any further questions.

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Question
Answer

Thank you for contacting us.

Unfortunately we have no data or experimental information to verify whether product ab52061 (anti-Cy5) cross reacts with AF-680.

Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

Thank you for your inquiry.

Unfortunately we have no information on whether this antibody cross-reacts with Cy5.5. However looking at a comparison of the molecules that I have attached I believe that it is probably highly unlikely that it will cross-react as the molecules are significantly different.

I checked our catalogue and we do not carry an anti-Cy5.5 antibody.

I am sorry that this information could not be more helpful. Please contact us with any other questions.

Read More

Answer

Thank you and your customer for your inquiry.

I am happy to confirm that ab52061 will only bind to Cy5 and not to the Rhodamine related Tamra or to biotin.

This is tested and guaranteed by our Abpromise.

I wish your customer good luck with their research.

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Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (brain)
Specification
brain

Mrs. Gitte Toft

Verified customer

Submitted Nov 04 2010

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

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