Goat F(ab) Anti-Rabbit IgG H&L (FITC) (ab7050)

Submit a review Q&A (2)References (3)

Overview

  • Product name
    Goat F(ab) Anti-Rabbit IgG H&L (FITC)
    See all IgG secondary antibodies
  • Host species
    Goat
  • Target species
    Rabbit
  • Specificity
    Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Fluorescein and anti-Goat Serum. No reaction was observed against anti-Pepsin or anti-Goat IgG F(c).
  • Tested applications
    Suitable for: ICC/IF, Immunomicroscopy, Flow Cyt, 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.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA

    BSA is IgG and protease free
  • Concentration information loading...
  • 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, pepsin digestion and chromatographic separation.
  • Conjugation notes
    Fluorescein isothiocyanate (FITC) (Molecular Weight 390 daltons) Absorption Wavelength: 495 nm Emission Wavelength: 528 nm Fluorochrome/Protein Ratio: 4.0 moles FITC per mole of Goat IgG Fab
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab7050 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
ICC/IF
Immunomicroscopy
Flow Cyt
ELISA
  • Application notes
    Flow Cyt: Use at an assay dependent dilution.
    IM: Use at an assay dependent dilution.

    Suitable for other antibody based fluorescent assays requiring extremely low background levels, absence of F(c) mediated binding, lot-to-lot consistency, high titer and specificity.

    Not tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.

    • References

    This product has been referenced in:
    • Scherz B  et al. mTh1 driven expression of hTDP-43 results in typical ALS/FTLD neuropathological symptoms. PLoS One 13:e0197674 (2018). Read more (PubMed: 29787578) »
    • Zhang J  et al. Hsp60 exerts a tumor suppressor function by inducing cell differentiation and inhibiting invasion in hepatocellular carcinoma. Oncotarget 7:68976-68989 (2016). IF . Read more (PubMed: 27677587) »
    See all 3 Publications for this product

    Publishing research using ab7050? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    I will attach few pictures here and in order to get your instruction further. MO3 and Med-2 is samples stained with AB53294/AB7050; and the Isotype-MBP and con-0.125 are stained with Purified Rabbit IgG Isotype /AB7050.

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    Answer

    Thank you for sending the ICC/IF pictures.

    The primary antibody stains as it should. The background between the cells can be reduced by exposing the cells for a shorter amount of time when taking an image orchanging the settings on your miscroscope. The images of the cells look very good in my opinion.

    What does the image med-2 show: primary + secondary or isotype control + secondary?
    This image looks like the primary antibody staining.

    The images with the isotype controls (Isotype-MBP and con-0.125) show too muchstainign for a negative control, and do not seem to be working correctly - as the isotype control is a negative control and should not show this staining intensity. How are Isotype-MBP and con-0.125 different from each other?

    In order to find out if the problem comes maybe from the secondary or isotype control, I'd suggest to do a no primary control (no primary and only secondary).This control should give youideallyno staining (or just weak non-specific background staining).
    Should this control work, thus, give no staining, then that would indicate that there isa problem with the isotype control.

    Since the primary antibody is a rabbit monoclonal, thus, using a rabbit monoclonal as isotype control would be recommended.
    We carry 2 such isotype controls in our catalog, should you be interested:
    ab125938 (https://www.abcam.com/Rabbit-IgG-monoclonal-SP137-isotype-control-ab125938.html)
    ab128142 (https://www.abcam.com/Rabbit-IgG-monoclonal-SP137-isotype-control-ab128142.html) prediluted

    Further, I'd suggest to use less secondary antibody (1/500 -1/1000) as this would help reducing the background you see between the cells as well.

    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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    Question

    ICC with human cell line
    isotype crtl: whole cell stained weakly, same as with primary Ab (no difference)
    primary Ab: 1/200 1.5 h
    block: 1% BSA
    2nd: 1/200 (ab7050)
    fix: acetone 5-10 min

    try no primary crtl, use less secondary (1/500 -1/1000)
    send images

    Read More
    Answer

    Thank you for contacting us.
    I am sorry to hear that one or both of these antibodies seem to not work very well.

    As we discussed over the phone, please send me the images with primary antibody as well as isotype control as this will help greatly to understand the problem better.

    Also, trying a no primary control, and/or using less secondary (1/500 -1/1000) could help reduce the non-specific background.

    I look forward to your reply and to assist you further.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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