Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)

Reviews (1)Q&A (1)References (75)

Overview

  • Product name

    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647)
    See all IgG secondary antibodies

  • Description

    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 647)

  • Host species

    Goat

  • Target species

    Rabbit

  • Specificity

    This antibody is specific to Rabbit IgG

  • Tested applications

    Suitable for: IHC-Fr, ICC/IF, ELISA, IHC-P, Flow Cytmore details

  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: 23% Glycerol, PBS, 1% BSA
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Purification notes

    This antibody was isolated by affinity chromatography using antigen coupled to agarose beads.

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • General notes

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab150079 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-Fr Use at an assay dependent concentration.
ICC/IF 1/200 - 1/1000.
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/2000.

Images

  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)
    Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)

    ICC/IF image of ab6046 in HeLa cells. The cells were 100% methanol fixed (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6046, 2µg/ml) overnight at +4°C. The secondary antibody ab150079 (shown in red) was used at 1µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

  • ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)
    ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)

    Cross-reactivity of the polyclonal secondary antibody ab182016 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. ab182016 was then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT.

    For the batch tested, ab182016 showed a cross-reactivity of 5-7% towards Human IgG and below 2% towards Mouse IgG, Rat IgG and Chicken IgY.

    This data was developed using the unconjugated antibody (ab182016).

  • ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)
    ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)

    Cross-reactivity of Goat anti-Rabbit IgG H&L (ab182016) and Goat anti-Rabbit IgG H&L obtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT. This data is from a representative dilution.

    This data was developed using the unconjugated antibody (ab182016).

  • Alexa Fluor® - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)
    Alexa Fluor® - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)

References

This product has been referenced in:

  • Liang C  et al. Y-box binding protein-1 promotes tumorigenesis and progression via the epidermal growth factor receptor/AKT pathway in spinal chordoma. Cancer Sci 110:166-179 (2019). Read more (PubMed: 30426615) »
  • He Y  et al. Dual-functional fluorescent molecular rotor for endoplasmic reticulum microviscosity imaging during reticulophagy. Chem Commun (Camb) 55:2453-2456 (2019). Read more (PubMed: 30734806) »
See all 75 Publications for this product

Publishing research using ab150079? 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

FOXA2 expressed in mouse E10.5 floor plate and notochord.

 Average
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
E10.5 mouse embryo section. Samples were fixed with PFA and stained before antigen retrieval with sodium citrate.

Primary: FOXA2 ab40874 (1:200).
Secondary: Alexa Fluor 647 ab150079 (1:1000).

Abcam user community

Verified customer

Submitted Jan 27 2014

Question

Product code: 150079
Inquiry: Dear Sir/Madam, I am using the products ab150079 and ab128251 to detect Cav3.2 in HEK cells expressing Cav3.2. However, in two attempts, I did not get any fluorescence in the Flow Cytometer, neither was any fluorescence visible in the fluorescent microscope. I use protocol from your website (https://www.abcam.com/index.html?pageconfig=resource&rid=11381). This protocol works brilliantly on the same cells but with other antibodies ab84815 and ab98692 (mouse monoclonal and anti-mouse FITC). In this case I got 40% staining of HEK/Cav3.2 cells with 2% non specific binding on WT HEK. The previous antibodies are rabbit and anti-rabbit. Do you have any idea what might be wrong? Is the first pair of antibodies not compatible or something like that? I look forward to receiving you reply. Yours sincerely,

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Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

- Have you used correct filters? The ab150079 absorption maxima is 650 nm while for FITC (ab98692) it is 495.
- Were the cells permeabilized?
- Is the sequence of insert protein is interpreted. It may well be that the insert sequence is missing which ab128251 binds.
The antibodies ab128251 and a84815 are raised against different part of the immunogen.
- Have you tried optimizing the results by incubating the antibody overnight or by increasing the concentration?

In order for checking the quality of ab150079 you may put a small drop of ab on slide and then seeing it under microscope. If it emits red colour then conjugate is fine and the problem may be with other steps. If it's not then problem is with secondary antibody.

It will be very helpful if you could provide the protcol followed step by step.

Please note that flow cytometry is not a tested application for ab128251 so the Abreview guarantee will not be valid. However if this antibody fails in ELISA, WB and ICC/IF then guarantee will apply as normal product.

Thank you very much for your cooperation. I will look forward to hearing from you soon.

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