Overview

  • Product name
    Goat Anti-Mouse IgG H&L (Cy5 ®) preadsorbed
    See all IgG secondary antibodies
  • Description
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Cy5 ®), pre-adsorbed
  • Host species
    Goat
  • Target species
    Mouse
  • Tested applications
    Suitable for: ELISA, ICC/IF, Flow Cyt, IHC-P, IHC-Frmore details
  • Minimal
    cross-reactivity

    Chicken, Cow, Goat, Guinea pig, Hamster, Horse, Human, Rabbit, Rat, Sheep more details
  • Immunogen

    Mouse IgG, whole molecule (H&L)

  • Conjugation
    Cy5 ®. Ex: 650nm, Em: 667nm

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities and extensive dialysis.
  • Conjugation notes
    Cy5.29 (Cyanine 5.29-OSu) (Molecular Weight 975 daltons) Absorption Wavelength: 650 nm Emission Wavelength: 667 nm Fluorochrome/Protein Ratio: 12.6 moles Cy5 per mole of Goat IgG
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • General notes
    This product or portions thereof is manufactured under license from Carnegie Mellon University under U.S. Patent Number 5,268,486 and related patents. Cy and CyDye are trademarks of GE Healthcare Limited. This material is also subject to proprietary rights of GE Healthcare Bio-Sciences Corp. and Carnegie Mellon University and made and sold under License from GE Healthcare Bio-Sciences Corp. This product is licensed for sale only for research. It is NOT licensed for any other use. There is no implied license hereunder for any commercial use. COMMERCIAL USE shall include: 1 Sale, lease, license or other transfer of the material or any material derived or produced from it. 2 Sale, lease, license or other grant of rights to use this material or any material derived or produced from it. 3 Use of this material to perform services for a fee for third parties. If you require a commercial license to use this material and do not have one, please return this material, unopened to Abcam Plc of 330 Cambridge Science Park, Cambridge, CB4 0FL, and any money paid for the material will be refunded.


    This secondary antibody is specifically designed for the detection of multiple primary antibodies (polyclonal or monoclonal) of different host species (human labelled with the fluorophore Cy5 here) in experiments where cells are simultaneously labeled without unwanted cross reaction.

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6563 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
ELISA 1/10000 - 1/50000. PubMed: 18691532
ICC/IF 1/1000 - 1/5000.
Flow Cyt Use at an assay dependent dilution.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Images

  • ab2792 (PDI antibody (RL90) - ER marker) staining human U2-OS (bone osteosarcoma) cells by ICC/IF. ab6563 (undiluted) was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Samy ZA  et al. Rat astrocytes during anoxia: Secretome profile of cytokines and chemokines. Brain Behav 8:e01013 (2018). Read more (PubMed: 29863786) »
  • Zalcman G  et al. Sustained CaMKII Delta Gene Expression Is Specifically Required for Long-Lasting Memories in Mice. Mol Neurobiol N/A:N/A (2018). Read more (PubMed: 29948945) »
See all 29 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Answer

Thank you for contacting us.

This antibody detects all mouse IgG, so it should work with both your IgG1 and IgG2 antibodies.

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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Abreviews
Application
Immunocytochemistry/ Immunofluorescence
This is a good functioning antibody with a stable conjugate.
The binding is very specific and the luminous intensity of the conjugate is good and also stable. We have used it in the ICC / IF with a 1/1000 Dilution.

Abcam user community

Verified customer

Submitted Oct 06 2015

Question

Hi,



I am trying to perform a modified sandwich ELISA/FluoSpot assay.

Basically what I want to do is coat wells in 96 or 384 well plate with an antibody (a mouse IgE,k) against protein X.

Then I wish to grow cells expressing protein X in the well.

Then after they have expressed some protein X, I wish to add a second antibody (a mouse IgG1,k) against protein X.

And finally a fluorescently conjugated secondary antibody against the second antibody (an anti mouse IgG1).

Then I will look in a microscope and identify cells producing large quantities of protein X and using a special technique I will purify those single cells.

Everything will preferably happen in cell media, preferably with serum, but if that is not feasible PBS can be used for the actual detection part (second antibody, secondary antibody, purification).



I am considering your “Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Cy5 ®), pre-adsorbed (ab6563)”.

Is there any chance this would cross react with the first antibody (mouse IgE)?

Is there any reason to believe this antibody would not work in the experiment above?

Also, I have to use a specific vendor for plates. They only supply “plastic” bottom plates. In your ELISPOT protocol you mention PVDF or nitrocellulose bottom plates. What is the point of the PVDF/nitrocellulose bottoms? I know that protein G can adhere to The “plastic” bottom, so I would expect antibodies would adhere as well or have I misunderstood something?



Kind Regards,

Read More
Answer

Thank you for your inquiry.

I used to perform many ELISPOTs by hand as you with plastic plates. The capture antibody was coated on the plates. The PVDF membrane bottom well plates areused when using a reader to analyze the results. The membranes are pushed through onto sticky tape and read in a reader with special software to identify spots.

I am not aware of a technique where the cells can be identified and sorted according to the production amount of protein X. I am assuming that protein X is secreted and therefore should be captured in the vicinity of the producing cell, but any slight movement of the plate will swirl the cells and make a identification impossible.

ab6563 is in general suitable as secondary to a mouse IgG1. But is is anti heavy and light chain. So it will pick up the light chain too. I suggest to choose an antibody specific to the heavy chain. I can recommend ab136127:

https://www.abcam.com/index.html?datasheet=136127 (or use the following: https://www.abcam.com/index.html?datasheet=136127).

This antibody is not specifically cross absorbed against IgE, but against Mouse IgM, IgG2a, IgG2b, IgG3 and IgA, pooled Human sera and purified Human paraproteins. ab136127 is tested in ELISA and therefore should also work in a ELISPOT setting as secondary.

I hope this information is helpful. Please do not hesitate to contact me again with any further questions in this matter.

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Answer

Sorry for the delay in getting back to you.

I can now confirm that the product has been cross adsorbed against the species listed as showing minimal cross reactivity. This includesBovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Rat, Rabbit or Sheep Serum Proteins.

I hope this information has been of help. If you have any further questions please do not hesitate to contact us again.

Read More

Answer

Thank you for contacting us and sorry for the delay in getting back to you in regards to your query.

I have looked into the antibody you are interested in, the Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Cy5 ®), pre-adsorbed (ab6563), and can share some more detail about its purification.This productis prepared by initially using immunoaffinity chromatography using Mouse IgG coupled to agarose beads. This is then followed by solid phase adsorptions to remove any unwanted reactivates to serum of other species and extensive dialysis.

Cross reactivity of this antibody has been assessed and noreactionhas beenobservedwith Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Rat, Rabbitor Sheep Serum Proteins.

I am checking further with the source of thisantibody to ascertain if the antibody has been pre-adsorbed with all of these species of serum and will get back to you as soon as I have this information.

I am sorry for the delay and any inconvenience this causes you.

Read More

Answer

Thank you for contacting Abcam.

All of the information regarding purification of the antibodies in our catalogue is available on our website. If we do not specify that Protein A/G was used in the purification process then they were not used to purify the antibody.

I have looked through the list you sent and for some there is no purification notes, as is the case for ab3354 as it is supplied as an Ascites fluid but for ab3356, it does say that Protein G was used for purification purposes.

Please let me know if there is anything else I can help you with.

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Answer

Thank you for your reply.

The information for how each antibody is purified is available on our website. Just go to the webpage for each antibody and the information is there. I have attached a document that shows you where to find that information.

If you have anymore questions after looking at the webpages, please let me know.

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Question
Answer

Thank you for contacting Abcam.

The antibody ab6563 was prepared from monospecific antiserum by immunoaffinity chromatography using Mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities and extensive dialysis.

Please let me know if there is anything else I can help you with.

Read More

Answer

Thank you very much for your interest in our antibodies. The difference between these two secondary antibodies it that ab6563 is pre-adsorbed against 10 species (chicken, cow, goat, guinea pig, hamster, horse, human, rabbit, rat, sheep), while ab97037 is pre-adsorbed against only 8 species (chicken, cow, goat, horse, human, rabbit, rat, sheep). It will depend on your lysate, which one would be better suited. If you are using a human cell line for example ab6563 would be better suited. Although we have not tested these antibodies, we believe they would work and could be used in Western blot, provided you have the appropriate equipment for the detection of Cy dyes. Please do not hesitate to contact us or the manufacturer of your detection system for more information. Given this, I can offer a discount off a future purchase if you buy ab6563 or ab97037 now, test it in Western blot and submit feedback to us. It doesn’t matter whether the feedback is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free secondary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab6563 or ab97037 in Western blot. I will then send a discount code. This code must be issued before purchasing ab so please wait for my reply before ordering. 2. Purchase ab6563 or ab97037 either by phone, fax, or online (www.abcam.com). 3. Test it in Western blot. 4. Let us know the results, positive or negative, preferably via an email to technical@abcam.com (images are great if you have them!) similar to our Abreview system (which can be found here: https://www.abcam.com/abreviews.) 5. After the feedback 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 secondary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the antibody turns out to be unsuitable for Western blot, you will still receive the discount on your next purchase after your feedback has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. Otherwise, if you are not interested sharing your results, i.e. in the above mentioned testing discount, please let me know and I could arrange for a discount when you place the order. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Question
Answer

Thank you for your phone call yesterday and for your questions about ab6563. I have been in touch with the lab, and I was told that small amounts of precipitate are common and should not affect the viability of the antibody. It is possible that the antibody was damaged if it went through a few freeze-thaw cycles, but it should be fine stored at 4C for couple weeks. If you

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1-10 of 15 Abreviews or Q&A

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