Overview

  • Product name
    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488)
    See all IgG secondary antibodies
  • Description
    Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)
  • Host species
    Donkey
  • Target species
    Rabbit
  • Specificity
    This antibody is specific to Rabbit IgG
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, IHC-P, ELISA, IHC-Frmore details
  • Immunogen

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

  • 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. Store In the Dark.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituents: 30% Glycerol, 1% BSA, PBS
  • 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 ab150073 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
ICC/IF 1/200 - 1/1000.
Flow Cyt 1/2000 - 1/4000.
IHC-P Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Images

  • ICC/IF image of ab6046 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal donkey 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, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab150073 Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) 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.

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10min).

  • ab104603 staining DYNLL1 in mouse cervical spinal cord tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked with 10% serum for 1 hour at 21°C. Samples were incubated with primary antibody (2 µg/ml in PBS/10% serum/0.1% Triton X-100) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG H&L (ab150073) (1/500) was used as the secondary antibody.

  • 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 donkey 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 Donkey anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150073) was used at 1/4000 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

This product has been referenced in:
  • Larasati YA  et al. Curcumin targets multiple enzymes involved in the ROS metabolic pathway to suppress tumor cell growth. Sci Rep 8:2039 (2018). Read more (PubMed: 29391517) »
  • Li C  et al. Nfatc1 Is a Functional Transcriptional Factor Mediating Nell-1-Induced Runx3 Upregulation in Chondrocytes. Int J Mol Sci 19:N/A (2018). Read more (PubMed: 29316655) »
See all 57 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abreviews
Application
Flow Cytometry
Sample: CAT1-overexpressed HCT116 cells
Condition: No fixation, No permeabilization
1st antibody: Rabbit anti-CAT1 antibody (ab37588) or Rabbit control IgG
2nd antibody: anti-Rabbit IgG Alexa-488 (ab150073)

Result: CAT1 was specifically detected by FACS.

Abcam user community

Verified customer

Submitted Aug 22 2017

Abreviews
Application
Immunohistochemistry (PFA fixed)
Works very well, I recommend

Yacine Tensaouti

Verified customer

Submitted Jul 26 2017

Application
Immunomicroscopy
Donkey anti-Rabbit IgG H&L (Alexa Fluor® 488) is nice to label the iNOS in mouse hepatocytes.

Abcam user community

Verified customer

Submitted Aug 07 2015

Abreviews
Application
Immunocytochemistry
This secondary marker was used at a dilution of 1/500 with PBS for 2 hours in combination with a primary H2Ax nuclear anti-body and the nuclear stainTOPRO3 in brain tissue from mice

Ms. Kirsty Ireland

Verified customer

Submitted Mar 18 2014

Application
Immunocytochemistry/ Immunofluorescence
Nice, bright clear staining seen using secondary antibody at a 1:200 dilution. Primary antibody used here was anti-Nanog (ab21624). Small amounts of speckled areas noted.

Dr. Sarah Ritson

Verified customer

Submitted Oct 07 2013

Abreviews
Application
Western blot
Application Western blot
Sample Mouse Rat lysate - Cytoplasm
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s)· Concentration: 5% · Temperature: 25°C
Other product details
Primary antibody- ab17820 Rabbit Polyclonal to Caspase 1
Dilution 1/1000, 1/2000. 1/5000
Incubation time 24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST
Secondary antibody
Application Western blot
Dilution 1/1000, 1/2000, 1/5000
Detection
Biorad Gel Doc XR+
Bands Specific: 45, 20 kDa
Stained membrane to check for protein transfer, all looked good. Incubated in both primary and secondary antibody and imaged, no bands were present at any of the dilutions.

Ms. Tammie Quinn

Verified customer

Submitted Sep 26 2013

Application
Immunocytochemistry/ Immunofluorescence
Following fixation in 4% PFA, the cells were assessed for Sox2 expression using 1:100 dilution of the primary antibody (ab97959) in 1% serum, 0.1% triton, 0.1% BSA in PBS, followed by detection using donkey polyclonal rabbit IgG Alexa 488 (ab150073) at 1:500. The results show that nuclear Sox-2 (green) was clearly observed. An isotype control IgG was run in parallel and showed no positive staining (not shown here). All nuclei were counterstained using DAPI (blue).

Abcam user community

Verified customer

Submitted Sep 02 2013

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

Sign up