Donkey Anti-Goat IgG H&L (Alexa Fluor® 405) preadsorbed (ab175665)

Overview

  • Product name
    Donkey Anti-Goat IgG H&L (Alexa Fluor® 405) preadsorbed
    See all IgG secondary antibodies
  • Host species
    Donkey
  • Target species
    Goat
  • Specificity
    By immunoelectrophoresis and ELISA this antibody reacts specifically with goat IgG and with light chains common to other goat immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Less than 1% cross reactivity to chicken, human, mouse, pig, rabbit and rat IgG was detected. This antibody may cross react with IgG from other species.
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, ELISA, IHC-P, Flow Cytmore details
  • Minimal
    cross-reactivity

    Chicken, Human, Mouse, Pig, Rabbit, Rat more details
  • Conjugation
    Alexa Fluor® 405. Ex: 402nm, Em: 421nm

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Please see notes section. Store In the Dark.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 30% Glycerol, 1% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • General notes

    We recommend storage time at 4°C should be minimal, since this may affect the signal strength.

    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 use products containing Alexa Fluor® dyes for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab175665 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.

We recommend the use of a dedicated 405 filter for optimal results not the DAPI filter. The DAPI filter may not excite until the maximum emission peaks of Alexa Fluor® 405 dye (see difference below) 
Ex max: Alexa Fluor® 405 = 402nm / DAPI = 359nm 
Em max: Alexa Fluor® 405 = 421nm / DAPI = 461nm

ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/2000.

Images

  • ICC/IF image of ab7291 stained 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 / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the primary antibody (ab7291, 1µg/ml) overnight at +4°C. Ab98800, goat anti-mouse IgG, was then added as a secondary bridging antibody, at 1/250 dilution for 1h. Ab175665 Alexa Fluor® 405 Donkey polyclonal Secondary Antibody to Goat IgG - H&L pre-adsorbed (blue) was then used at 1µg/ml for 1h as a tertiary antibody. DRAQ5™ (ab108410) was used to stain the cell nuclei (red) at a concentration of 1.25µM.

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

  • Postnatal day 6 mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% BSA in 0.1% Triton X-100 + PBS. Sections were incubated with either (A) no primary antibody or (B) Goat Anti-DDX4 at a 1/800 dilution for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Donkey Anti-Goat IgG H&L (Alexa Fluor® 405) (ab175665) applied at a 1/500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.

References

ab175665 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abreviews
Application
Immunohistochemistry (Frozen sections)
This antibody works well at a 1:500 dilution and gives a bright signal with little discernible nonspecific staining.
Postnatal day 6 mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% BSA in 0.1% Triton X-100 + PBS. Sections were incubated with either (A) no primary antibody or (B) Goat Anti-DDX4 at a 1:800 dilution for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Donkey Anti-Goat 405 (abcam ab175665) applied at a 1:500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.
Username

Mr. Bryan Niedenberger

Verified customer

Submitted May 06 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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