Overview

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

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

    Rabbit IgG, whole molecule

  • 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 Rabbit 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: 6.0 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 secondaryantibody is specifically designed for the detection of multiple primary antibodies (polyclonal or monoclonal) of different host species in experiments where cells are simultaneously labeled without unwanted cross reaction.

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6564 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
Flow Cyt Use at an assay dependent dilution.
ICC/IF 1/1000 - 1/5000.
ELISA 1/10000 - 1/50000.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Images

  • Immunofluorescence stainings of human prostate tissues

    Sections were double labeled for different PAK isoforms, together with calponin (marker for smooth muscle cells), pan-cytokeratin (marker for epithelial cells of glands), or tyrosine hydroxylase (TH, marker for catecholaminergic nerves). Yellow color in merged pictures may indicate colocalization of targets. Shown are representative stainings from series with tissues from n = 5 patients for each combination.

    ab6908 was used as secondary antibody to visualize various rabbit anti-PAK primary antibodies (red).

    (From Figure 2 of Hennenberg et al)

  • ab11175 staining Histone H2A.X in mouse testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked with 4% BSA for 2 hours at 37°C. Antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in PBS/4% BSA/0.05% Triton X-100) for 48 hours at 4°C. A Cy5®-conjugated goat anti-rabbit IgG H&L preadsorbed (ab6564) (1/300) was used as the secondary antibody.

    See Abreview

  • Ab6564 was used at dilution 1/300 with the primary antibody ab19898 in Flow Cyt. See the review on ab19898.
  • Ab6564 was used at dilution 1/500 with the primary antibody ab24586 in ICC/IF. See the review on ab24586.

References

This product has been referenced in:
  • Yu Q  et al. Inhibition of prostatic smooth muscle contraction by the inhibitor of G protein-coupled receptor kinase 2/3, CMPD101. Eur J Pharmacol 831:9-19 (2018). IF . Read more (PubMed: 29698717) »
  • Azizi F  et al. The combined effects of three-dimensional cell culture and natural tissue extract on neural differentiation of P19 embryonal carcinoma stem cells. J Tissue Eng Regen Med 12:1909-1924 (2018). Read more (PubMed: 29905008) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Answer

Thank you for contacting us.

Both of these antibodies will be guaranteed for your purposes, and we would expect them to give equivalent signal and specificity. The main difference between the two is the recommended dilutions for ICC/IF. You should be able to get more tests per vial by using ab6564.

I hope this helps, please let me know if you need any additional information or assistance.

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Question
Answer

Thank you for your reply.

They could try the secondary ab6564 at 1/300 - 1/500.

https://www.abcam.com/Rabbit-IgG-secondary-antibody-H-L-ab6564.html#IgG-Secondary-antibodies-ab6564-1.jpg

I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for your inquiry.

We recommend using this antibody in FC 1"10 - 1:100 dilution. We tested permeabilized HeLa cells and the histograph can be found on the datasheet: https://www.abcam.com/Desmin-antibody-Y266-ab32362.html

This is the recommended protocol:

Flow Cytometry Protocol for Intracellular Staining

1. Solutions and Reagents

1.1. 1X PBS

1.2. Blocking buffer: 0.5% BSA in 1X PBS

1.3. 2% paraformaldehyde (1% solution - optional for storing samples)

1.4. 1X FACS permeabilizing solution (BD Biosciences cat. #340973)

1.5. Fluorescently-conjugated secondary antibody (various forms)

2. Protocol

2.1. Collect 1x106 cells/sample.

2.2. Wash cells once with blocking buffer.

2.3. Fix cells with 2% paraformaldehyde and incubate at room temperature for 10 min.

2.4. Wash cells once with blocking buffer.

2.5. Add 0.5 ml 1X FACS permeabilizing solution and incubate at room temperature for 10 min.

2.6. Wash cells once with blocking buffer.

2.7. Incubate cells in blocking buffer for 30 min at room temperature.

2.8. Add primary antibody at the appropriate dilution and incubate for 30 min at room temperature.

2.9. Wash twice with blocking buffer and incubate with fluorescently-conjugated secondary antibody for 30 min at room temperature.

2.10. Wash cells twice with blocking buffer.

2.11. Re-suspend cells in 1X PBS and analyze on flow cytometry. Samples can be kept in 1% paraformaldehyde at 4 °C overnight.

I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for your email.

I can confirm that ab6564 is not yet tested experimentally inWB and therefore we cannot guarantee that ab6564 works in WB. But it is very likely to work in WB.

As my colleague who is not in the office today explained: the testing discount program requires you to purchase the antibody that you would like to test (here in WB). After submission of the Abreview the code will be valid for any product from our catalog.

Even if the tested antibody is not suitable for WB you will receive a free product of your choice to the value of the tested antibody.

You will also receive a free product of your choice from our catalog if the tested antibody works.

Unfortunately, we do not have any anti rabbit secondary antibodies that are conjugated to Cy5 and are tested in WB in our catalog at this moment in time.

I hope I could prevent any misunderstandings and hope you will decide for the testing program. I wish you good luck for your research.

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Answer

Thank you for your reply.

Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

As I pointed out before, the antibody is to be used in an untested application, you are eligible for our testing discount program, which will leave you with two antibodies for the price of one. We think thisis a fair offer as you will get a reward even if the antibody is not suitable for you.

We always appreciate customer feedback, whether positive or negative, whether from a testing discount or from the customers initiative, and we make all product information available to researchers. In addition, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates. This includes the testing discount reviews as well.


I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Answer

Thank you very much for your interest in ab6564 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Cy5 ®), pre-adsorbed.

To our knowledge, ab6564has not been tested in WB, and so has neither of our other secondary antibodies. Therefore, I can offer a discount off a future purchase if you buy ab6564 now, test it in WB 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: 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 ab6564 inWB. I will then send a discount code. This code must be issued before purchasing ab6564 so please wait for my reply before ordering.

2. Purchase ab6564 either by phone, fax, or online (www.abcam.com).

3. Test it in WB.

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 secondary antibody 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 ab6564 turns out to be unsuitable for WB, 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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Answer

Thank you very much for your interest in Abcam and our products..

To our knowledge,the following antibodies you listed have not been tested in IHC-P. Tested applications and species are listed on the individual datasheets. These are updated as soon as we receive any new information.

ab25644
ab24941
ab86900
ab41467,
ab60077,
ab25175,
ab24985,
ab59437,
ab116223

Therefore, I can offer a discount off a future purchase if you buythese untested antibodies now, test in IHC-P 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.One discount code would be provided for each individualantibody thatwould be to the value of 1 free primary 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.If you could let me know whichparticular antibodies youwant to test. I will then send a discount code for each of these antibody. This code must be issued before purchasing the antibodiesso please wait for my reply before ordering.

2. Purchase the antibodies either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-P.

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 primary 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 any of the antibodiesturns out to be unsuitable for IHC-P, 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.

The following antibodies you listed have already been tested in IHC-P, and so we are not able to provide a testing discount for these on this occasion. As they have been tested in IHC-P, we can guarantee them for fluorescent studies on paraffin embedded tissue as well.

ab112470
ab61219
ab6564

We do not charge individual shipping for each item. We would charge one shipping fee for eachorder. Extra shipping charges are applied if there is a bulky item such as a kit, or a dry ice item on the order.

With regards to discounts for new laboratories, I have spoken to our marketing department and believe thatone ofyour colleagues, Dr. ##### has already been in touch with us. A 25% discount has been arranged for orders from his laboratory for the next 1 month. Could you confirm that you are working in the same laboratories? I hope this discount will prove helpful in setting up the new laboratory. As we have already arranged these 2 types of discount, I am sorry we are not able to offer a further bulk discount or shipping discount on this occasion.

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

Read More

Answer

Yes, the secondary antibody to Rabbit IgG - H&L (Cy5 ®) ab6564 is compatible with the anti-SAP102 antibody ab83980.

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Question
Answer

Thank you for your enquiry. Cy5 emits the color red (it is located at the Far-Red end of the spectrum). The following link on our website should be helpful for you (there is a Fluorochrome table on that particulat page): http://abcam2/index.html?pageconfig=tech&mode=choosing So, if Cy5 overlaps with Fitc, the color seen would be yellow.

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Answer

Thank you for your email. Ab6075 has been tested for application in IHC-Frozen sections (IHC-Fr). It has not yet been tested for application in IHC-formalin fixed paraffin embedded tissue sections. For ab6564, Goat polyclonal to Rabbit IgG H&L (Cy5), H&L stands for heavy and light chain. If you have any more questions, please contact us again.

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1-10 of 11 Abreviews or Q&A

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