Goat Anti-Rabbit IgG H&L (FITC) (ab6717)

Reviews (1)Q&A (22)References (185)

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

  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (FITC) (ab6717)
    Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (FITC) (ab6717)Kumar S et al. The role of tumour suppressor PDCD4 in beta cell death in hypoxia. PLoS One 12:e0181235 (2017). Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Subcellular localisation of PDCD4 was assessed by immunocytochemical staining. ARIP cells were seeded at 3×105 cells/well on sterile cover slips in six well plates and incubated overnight under standard culture conditions.

    Following specific experimental conditions, media was removed and cells were washed with PBS and fixed with 3.7% formalin. Cells were then permeabilised with 0.1% triton X-100 in PBS followed by blocking in blocking buffer (10% Goat serum; 2% BSA; 0.2% Tween 20; 0.7% Glycerol in PBS) for 1 hour. Cells were incubated overnight with ab51495 at 1/100 dilution followed by ab6717 (green) at 1/80 dilution. The nuclear counterstain is DAPI (Blue).

    Results are representative of three separate experiments and images were representative of six separate fields. PDCD4 localized and expressing At G0: Cytoplasmic and very low expression; At N12: Cytoplasmic and low expression; At N24: very high cytoplasmic and low nuclear expression; At H12: Cytoplasmic expression and At H24 very high cytoplasmic and low nuclear expression. Over-all PDCD4 was highly expressed in the cytoplasm, with very low expression under serum starved conditions

  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (FITC) (ab6717)
    Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (FITC) (ab6717)This image is courtesy of an anonymous Abreview
    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (FITC) (ab6717)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (FITC) (ab6717)
    Ab6717 was used at dilution 1/200 with the primary antibody ab36823 in IHC-P. See the review on ab36823.
  • Flow Cytometry - Goat Anti-Rabbit IgG H&L (FITC) (ab6717)
    Flow Cytometry - Goat Anti-Rabbit IgG H&L (FITC) (ab6717)
    Ab6717 was used with the primary antibody ab66506 in Flow Cyt. See Abreview on ab66506.

References

This product has been referenced in:

  • Chignola R  et al. Collective radioresistance of T47D breast carcinoma cells is mediated by a Syncytin-1 homologous protein. PLoS One 14:e0206713 (2019). Read more (PubMed: 30699112) »
  • Zhang R  et al. Mechanical strain triggers differentiation of dental mesenchymal stem cells by activating osteogenesis-specific biomarkers expression. Am J Transl Res 11:233-244 (2019). Read more (PubMed: 30787982) »
See all 185 Publications for this product

Publishing research using ab6717? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 23 Abreviews or Q&A

Question

Inquiry: Sehr geehrte Damen und Herren, ich interessiere mich für die folgenden Produkte:
Anti-Osteopontin antibody (ab8448)
Anti-TRAF6 antibody (ab94769)
Anti-TGF beta 1 antibody [TB21] (ab27969)
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC) (ab6717)
Könnten Sie mir bitte ein konkurrenzfähiges Angebot für diese Antikörper schicken? Mit freundlichen Grüßen

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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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Vinculin Staining

 Good
Abreviews
abreview image
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

Question

Hi,

Thanks for the info! I will have a good read through. Just one quick question for you:

My cells are already fixed in 3:1 Methanol:Acetic acid. Does this mean I can skip permeabilisation as there is methanol in this?

Thanks again,

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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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Question

Hi,

Thanks for the info! I will have a good read through. Just one quick question for you:

My cells are already fixed in 3:1 Methanol:Acetic acid. Does this mean I can skip permeabilisation as there is methanol in this?

Thanks again,

Read More
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

Wants to perform IF. Asked what diluent to use.

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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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Question

We have some question about Goat polyclonal Secondary Antibody to Rabbit IgG ab6717.
Recommended dilution-1\1000
how much diluent we must add to vial with 1mg (2mg\ml) ?

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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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Question

Could you please tell me the the isotype of your product "Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC) (ab6717)". I am looking for an isotype control for ab6717 for FACS.

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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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Question

I have question which of negative control could you recommend for products: ab112538, ab6717, ab97050.

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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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Question

Dear Sir or Madam,



I would like to know if it is possible to obtain a free sample of the following antibodies: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC) (ab6717).



I would appreciate a reply at your earliest convenience.



Yours faithfully,

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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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Question

We’ve packaged the other antibody up and are awaiting FedEx pickup.

Thanks for sending the correct antibody asap.

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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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