Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 555) preadsorbed (ab150062)

Overview

  • Product name
    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 555) preadsorbed
    See all IgG secondary antibodies
  • Description
    Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 555), pre-adsorbed
  • 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, ICC/IF, ELISA, Flow Cyt, IHC-Pmore details
  • Minimal
    cross-reactivity

    Human, Mouse, Rat more details
  • Conjugation
    Alexa Fluor® 555. Ex: 555nm, Em: 565nm

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. 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 human, mouse 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

    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 ab150062 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.
Flow Cyt 1/2000.
IHC-P Use at an assay dependent concentration.

Images

  • ICC/IF image of ab6046 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal donkey serum / 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 antibody (ab6046, 1µg/ml) overnight at +4°C. The secondary antibody (yellow) was ab150062 Alexa Fluor® 555 donkey anti-rabbit IgG (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • 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 ) anti-DDX4 (ab13840) for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Goat-Anti Rabbit 555 (ab150062) applied at a 1/500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS. 

  • HeLa cells showing negative staining by ICC/IF using only secondary antibody. The cells were 100% methanol fixed (5 min) and then incubated in 1% BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The secondary antibody (yellow) was ab150062 Alexa Fluor® 555 donkey anti-rabbit IgG (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Paliwal D  et al. Hepatitis E Virus (HEV) egress: Role of BST2 (Tetherin) and interferon induced long non- coding RNA (lncRNA) BISPR. PLoS One 12:e0187334 (2017). Read more (PubMed: 29091957) »
  • Rauber S  et al. Resolution of inflammation by interleukin-9-producing type 2 innate lymphoid cells. Nat Med 23:938-944 (2017). IF . Read more (PubMed: 28714991) »

See all 3 Publications for this product

Customer reviews and Q&As

Abreviews
Application
Immunohistochemistry (Frozen sections)
I have used this antibody at a 1:500 dilution with many different primary antibodies. It works well and and gives a bright signal with no 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 ) anti-DDX4 (Abcam ab13840) for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Goat-Anti Rabbit 555 (Abcam ab150062) 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 Mar 29 2016

Application
Immunocytochemistry/ Immunofluorescence
iPS derived neurons (35 days) were fixed in 4% formaldehyde (10 min) and then incubated in 1% serum, 0.1% triton, 0.1% BSA in PBS for 30 mins to permeabilise the cells and block non-specific binding. The cells were then dual stained (simultaneous method) with antibody ab18207 (BIII Tubulin at 1/100 dilution) and Vglut1 (green) for 1h at room temp. Secondary antibodies used were Donkey anti-Rabbit IgG H&L Alexa 555 (ab150062) (red) and donkey anti mouse IgG 488 at 1:250 dilution and incubated for 1h at room temp. Total cells were stained with DAPI (blue). Positive BIII-Tubulin staining was observed in processes (green).
Username

Abcam user community

Verified customer

Submitted Aug 15 2014

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

Sign up