Overview

  • Product name
    Goat Anti-Rabbit IgG H&L (FITC)
    See all IgG secondary antibodies
  • Host species
    Goat
  • Target species
    Rabbit
  • Tested applications
    Suitable for: IHC-FoFr, Immunomicroscopy, Flow Cyt, IHC-P, IHC-Fr, ICC/IF, ELISAmore details
  • Immunogen

    Rabbit IgG whole molecule

  • Conjugation
    FITC. Ex: 493nm, Em: 528nm

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C. Store In the Dark.
  • 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
    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.
  • Conjugation notes
    Fluorescein isothiocyanate (FITC) (Molecular Weight 390 daltons) Absorption Wavelength: 495 nm Emission Wavelength: 528 nm Fluorochrome/Protein Ratio: 2.7 moles FITC per mole of Goat IgG
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab6717 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-FoFr Use at an assay dependent concentration. PubMed: 24216136
Immunomicroscopy Use at an assay dependent concentration.
Flow Cyt 1/500 - 1/2500.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/1000 - 1/5000.
ELISA 1/10000 - 1/50000.

Images

  • ab11320 (Rabbit polyclonal to gamma Tubulin - Centrosome Marker) at 1/400 dilution staining human neuroblastoma cells by ICC/IF. The cells were fixed with paraformaldehyde and blocked with serum and then incubated with the primary antibody for 16 hours. ab6717 was used as the secondary.

    See Abreview

  • Ab6717 was used at dilution 1/200 with the primary antibody ab36823 in IHC-P. See the review on ab36823.
  • Ab6717 was used with the primary antibody ab66506 in Flow Cyt. See Abreview on ab66506.

References

This product has been referenced in:
  • Gao J  et al. In vitro investigation of the mechanism underlying the effect of ginsenoside on the proliferation and differentiation of neural stem cells subjected to oxygen-glucose deprivation/reperfusion. Int J Mol Med 41:353-363 (2018). Read more (PubMed: 29138802) »
  • Mohamed BM  et al. Induction of protein citrullination and auto-antibodies production in murine exposed to nickel nanomaterials. Sci Rep 8:679 (2018). Read more (PubMed: 29330439) »
See all 136 Publications for this product

Customer reviews and Q&As

1-10 of 23 Abreviews or Q&A

Answer

Vielen Dank für Ihre Anfrage.

Ich freue mich ihnen mitteilen zu koennen dass wir fuer den gesamten November unsere Aktion 4 fuer 3 laufen haben. Bei der Bestellung von 4 Produkten bekommen Sie das guenstigste Produkt kostenfrei.

Sie koennen ganz einfach per E-mail bestellen: mailto:orders@abcam.com


Geben sie bei der Bestellung bitte den Aktionscode an: 4-FOR-3-EB3AB


Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Mouse Osteoblast cells were grown on cover slips for 48 hrs. Then Cells were fixed with formaldehyde vapor for 10 mins. Cells were permeabilized with Triton-X 100 for 5-7mins. 3% BSA was used for blocking (1hr). 1:100 dilution of rabbit anti-vinculin antibody to mouse was added onto the cells and incubated for 1hr. Then 1:1000 dilution of anti-rabbit FITC conjugated secondary antibody was added on to the cells and incubated for 1hr. Before proceeding to each step cells were washed with PBS.

Abcam user community

Verified customer

Submitted Jul 05 2013

Answer

I am sorry for the delay in getting back to you. I now have further information to share in regards to how the Anti-Histone H2B antibody [EP957Y] (ab52599) has been used in staining of fixed cells.

The image presented on the datasheet of ab52599 shows the results obtained when using the antibody in the staining of Histone H2B in HeLa cells. This was achieved using acetone at -20°C for 5 minutes to fix and permeabilize the cells. The antibody has also been used successfully with fixation using 2% paraformaldehyde for 20 min and permeabilization with 0.1% Triton-X100 for 5 min. This is described in more detail in the following protocol:

http://www.epitomics.com/products/support/icc

I hope this information has been of help. I also wanted to bring to your attention a webinar we are hosting this afternoon on "Optimizing IHC/ICC results through careful experimental design" which may be of interest to you. More information on this can be found from the following link. It is free to attend.

https://www.abcam.com/index.html?pageconfig=resource&rid=15358

If you have any further questions please do not hesitate to ask.

Until then, I wish you all the best with your research.

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Answer

Thanks for your reply. I am glad you found the information useful.

With the fixative you intend to use I do not think you will require permiabilisation. However, methanol is sometimes not suitable for some targets/antibodies. It may therefore need some optimisation in order to obtain the best results.

I am currently contacting the lab to see how this antibody was used in IF previously and what fixation was used. Once I have further information to share I will get back to you.

In the meantime, if you have any further questions, please do not hesitate to ask.

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Question
Answer

Thank you for contacting us this morning in regards to the anti-Histone H2B antibody (ab52599) and the Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC) (ab6717).

These antibodies should work fine in combination for the experiment you are currently planning. As discussed over the phone, we have quite a useful protocol guide which can be accessed from the following link:

https://www.abcam.com/index.html?pageconfig=resource&rid=13111

You can also request a hard copy of this from the link. This provides a good overview of the main antibody applications as well as providing troubleshooting should the results not be as expected. As you are working with a nuclear target, I would suggest introducing a permiabilisation step. For example incubating your samples with 0.2% triton for 10 minutes (unless you are using acetone or methanol for fixation). The antibody could be diluted in PBS with 0.1% tween as well as 0.1-1% BSA. We would normally recommend using overnight incubation at 4°C as this tends to result in a more specific signal.

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

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Answer



In order to dilute an antibody 1\1000, you can, for example, add 1ul for every 1000ul of diluent.

Regarding your question, If the vial of ab6717 contains 1mg at 2mg/ml, then it contains about 0.5ml of volume. To diute the entire 0.5ml of ab6717, you would dilute the contents of the vial (0.5ml) into a final volume of 500mls, again a 1\1000 dilution.

Please note the unused portion of the antibody should not be diluted and stored. The unsued portion should be aliquoted (at the original concentration) and stored at -20 degrees C.

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Answer

Thank you for contacting us.

We have an isotype control of goat IgG, conjugated to FITC. The catalogue number is ab37374.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=37374).

This is sold as "100 tests" (10ul per test). The concentration is 0.1mg/ml, and the volume is 1ml.

Please note that ab6717 has a different concentration (2mg/ml) and also a different concentration of the preservative sodium azide (0.01% as compared to 0.1% for ab37374, 10-fold higher). This may be an important consideration if you are staining live cells, as sodium azide can be cytotoxic.

Please let me know if you have any questions.

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Answer

Thank you for contacting us.
According to the datasheet, the this protein is not expressed in most lymphoid cell lines: B-cells, T-cells and NK cells. Therefore, I would suggest to use a T-cell line like Jurkat lysate as a negative control.

https://www.abcam.com/index.html?datasheet=7899 https://www.abcam.com/index.html?datasheet=7899.

As a "negative control" for WB I woul suggest to perform a non-primary control, which simply means skipping the promary antibody incubation.

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

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

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 for your reply and for returning the vial of ab6785.

The vial of ab6717 will be arriving tomorrow on the order ***.


Please let me know if you need anything else, and have a wonderful evening.

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1-10 of 23 Abreviews or Q&A

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