Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) preadsorbed (ab175649)

Overview

  • Product name
    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) preadsorbed
    See all IgG secondary antibodies
  • Host species
    Donkey
  • Target species
    Rabbit
  • Specificity
    By immunoelectrophoresis and ELISA this antibody reacts specifically with rabbit IgG and with light chains common to other rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Less than 0.1% cross reactivity to bovine, chicken, goat, human, mouse, pig and rat IgG was detected. This antibody may cross react with IgG from other species.
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, ELISA, ICC/IF, Flow Cytmore details
  • Minimal
    cross-reactivity

    Chicken, Cow, Goat, Human, Mouse, Pig, Rat more details
  • Immunogen

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

  • 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
  • Purification notes
    Antiserum was cross adsorbed using bovine, chicken, goat, human, mouse, pig and rat immunosorbents to remove cross reactive Antibodies. The antibody to rabbit IgG was isolated by affinity chromatography using antigen coupled to agarose beads.
  • 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 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 ab175649 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.
IHC-P Use at an assay dependent concentration.
ELISA 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

Flow Cyt 1/2000.

Images

  • ICC/IF image of ab6046 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the primary antibody (ab6046, 1µg/ml) overnight at +4°C. The secondary antibody (blue) was ab175649, Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) preadsorbed, used at 2µg/ml for 1h at room temperature. DRAQ5™ (ab108410) was used to stain the cell nuclei (red) at a dilution of 1/4000.

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

References

This product has been referenced in:
  • McWilliams TG  et al. Basal Mitophagy Occurs Independently of PINK1 in Mouse Tissues of High Metabolic Demand. Cell Metab 27:439-449.e5 (2018). Read more (PubMed: 29337137) »
  • Koba S  et al. Sympathoexcitation by hypothalamic paraventricular nucleus neurons projecting to the rostral ventrolateral medulla. J Physiol N/A:N/A (2018). Read more (PubMed: 30019338) »
See all 8 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry
Application: Fluorescence IHC staining of mouse brain tissue for tyrosine hydroxylase in 4% paraformaldehyde-fixed free-floating coronal 20 uM sections

Methods:
The sections were washed with phosphate buffer (PB) followed by 0.05 M tris-buffered saline containing 1% Triton-X (TBS-TX, pH 7.4). The sections were blocked with 5% bovine serum albumin for 1 h and incubated with 1:2000 TH (rabbit monoclonal) in TBS-TX overnight. Sections were washed with PB-TX and incubated with secondary antibody Alexa Fluor 405 donkey anti-rabbit IgG (Abcam, ab175649) for 1 h at room temperature. After rinsing with PB, sections were mounted on glass slides with Prolong Gold antifade reagent.

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Verified customer

Submitted Aug 03 2015

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

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