Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

Overview

  • Product name
    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed
    See all IgG secondary antibodies
  • Description
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed
  • 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, horse, 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, Flow Cyt, IHC-P, ELISAmore details
  • Minimal
    cross-reactivity

    Human, Mouse, Rat more details
  • Conjugation
    Alexa Fluor® 488. Ex: 495nm, Em: 519nm

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: PBS, 30% Glycerol, 1% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Antiserum was cross adsorbed using a human, mouse and rat immunosorbents to remove cross reactive antibodies. 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 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

Associated products

Applications

Our Abpromise guarantee covers the use of ab150081 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.
Flow Cyt 1/2000 - 1/4000.

ab199091-Rabbit monoclonal IgG (Alexa Fluor® 488), is suitable for use as an isotype control to complement this secondary antibody.

IHC-P Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed images

  • ICC/IF image of ab8227 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat 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 (ab8227, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab150081 Alexa Fluor® 488 goat 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.

    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 ) 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 488 (ab150081) applied at a 1/500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.

  • Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

    Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)

    Preparation:

    Fix in 3%PFA in PBS for 30 min at RTIncubate in 7.5% sucrose-PBS for 3h at RTIncubate in 15% sucrose-PBS at 4 degree Celsius overnightEmbed the EBs in tissue-Tek OCT compoundCut frozen sections to 4-20 µm thickness

    Primary antibody 1: Rabbit anti cytokeratin 8 (ab53280), 1:100

    Primary antibody 2: Rat anti-perlecan, 1:100 
    Secondary antibody 1: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200

    Secondary antibody 2: Goat polyclonal Secondary Antibody to Rat IgG - H&L (Cy5®) pre-adsorbed, 1:200
    Nuclei were counterstained with DAPI 

  • Overlay histogram showing Jurkat cells stained with ab16669 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

References for Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

This product has been referenced in:
  • Sarenac T  et al. Single-cell analysis reveals IGF-1 potentiation of inhibition of the TGF-ß/Smad pathway of fibrosis in human keratocytes in vitro. Sci Rep 6:34373 (2016). Read more (PubMed: 27687492) »
  • Jones-Paris CR  et al. Basement membrane ultrastructure and component localization data from uterine tissues during early mouse pregnancy. Data Brief 9:931-939 (2016). IHC-Fr . Read more (PubMed: 27896299) »

See all 5 Publications for this product

Product Wall

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 488 (Abcam ab150081) 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 09 2016

Application
Immunocytochemistry/ Immunofluorescence
This antibody has been used extensively with a number of different primary antibodies generated in rabbits. It performs extremely well at a 1:200 dilution.
Attached is an immunofluorescent image of MeOH-fixed HeLa cells that were immunofluorescently labeled with a Lamin A/C (nuclear membrane) antibody (ab133256). The secondary antibody has been pseudo-colored green in the merged image. DNA was counterstained with DAPI.
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Mar 28 2014

Application
Immunohistochemistry
Application: Immunohistochemistry (PFA perfusion fixed frozen sections) or IHC(FoFr)

I used this antibody on PFA fixed, sucrose cryoprotected frozen sections from the mouse brain (below is a brief description). The primary a.b. I used was anti-mGluR5 (ab53090), which was somewhat difficult to get working.

Full Description:
Transcardially perfused the mouse with 4% PFA. Dissected the brain and placed in PFA for a few hours later, placed in 30% sucrose overnight for cryoprotection. Then, snap/flash froze the brain and used a cryostat for sectioning into 20 micron coronal slices. Applied primary anti-mGluR5 (ab53090) antibody, followed by the secondary antibody (dilution: 1/200), according to standard protocols. No blocking, no antigen-retrieval, and no permeabilization was done.
Username

Abcam user community

Verified customer

Submitted Jan 29 2014

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

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