Overview

  • Product nameGoat Anti-Rabbit IgG H&L (HRP)
    See all IgG secondary antibodies
  • Target speciesRabbit
  • Tested applicationsIHC-P, WB, ELISA, Immunomicroscopy, Dot Blot, ICC/IF, IHC-Frmore details
  • Immunogen

    Rabbit IgG, whole molecule

  • ConjugationHRP

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C.
  • Storage bufferPreservative: 0.01% Gentamicin Sulfate
    Constituents: 10mg/ml BSA, 0.15M Sodium chloride, 0.02M Potassium phosphate, pH 7.2
  • Concentration information loading...
  • PurityIgG fraction
  • Purification notesThis product was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit IgG coupled to agarose beads.
  • Conjugation notesHorseradish Peroxidase (HRP)
  • ClonalityPolyclonal
  • IsotypeIgG
  • General notesMany of our customers have reported seeing brown precipitates in the vials. The brown precipitates are very common with HRP conjugated antibodies; we suggest vortexing the vial and using this antibody as normal. Our customer’s feedback says the antibody worked great. If in case the antibody fails to give results then please contact our Scientific Support team for assistance.
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab6721 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-P 1/1000.
WB 1/2000 - 1/20000. Suggested working dilution of 1/3000 (see PMID: 17222046). In addition, found to work at 1/20000 (see PMID: 16936283).
Working dilutions are highlighted in the table below. Please note that the antibody can be diluted to 1:48,000 to 1:207,000 in many instances.
ELISA 1/120000.
Immunomicroscopy Use at an assay dependent dilution.
Dot Blot Use at an assay dependent dilution.
ICC/IF 1/1000 - 1/5000.
IHC-Fr 1/1000.

Goat Anti-Rabbit IgG H&L (HRP) images


  • developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 10 seconds

    This image is courtesy of an Abreview submitted by Maryna Polyakova.

    See Abreview


  • developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview.

    See Abreview

  • ab5095 Immunoprecipitating RNA polymerase II in human K562 whole cell lysate. 20µg of cell lysate was incubated with primary antibody (1/1000 in TBST) and matrix (Protein A) for 12 hours at 4°C. For western blotting a HRP-conjugated goat anti-rabbit IgG H&L (ab6720) (1/1000) was used to confirm successful immunoprecipation.

    See Abreview

  • ab3580 staining glucocorticoid receptor in mouse epididymis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with bouin's solution and blocked with 1.5% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1000) for 14 hours at 4°C. A HRP-conjugated goat anti-rabbit IgG H&L (ab6720) (1/200) was used as the secondary antibody.

    See Abreview

  • Ab6721 was used at dilution 1/100 with the primary antibody ab11370 in IHC-P. See the review on ab11370.
  • Ab6721 was used at dilution 1/100 with the primary antibody ab35604 in IHC-Fr. See the review on ab35604.

References for Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

This product has been referenced in:
  • Gasperin BG  et al. Expression of receptors for BMP15 is differentially regulated in dominant and subordinate follicles during follicle deviation in cattle. Anim Reprod Sci 144:72-8 (2014). Read more (PubMed: 24388700) »
  • Westerbeck JW  et al. A SUMO-targeted ubiquitin ligase is involved in the degradation of the nuclear pool of the SUMO E3 ligase Siz1. Mol Biol Cell 25:1-16 (2014). WB . Read more (PubMed: 24196836) »

See all 92 Publications for this product

Product Wall

Application Western blot
Experimental conditions:
Bone tissue lysate (tibia) extracted with RIPA buffer
10% SDS-PAGE
Blocking 1h with 5% milk in TBS-T at RT
Incubation with primary AB 1:1000 overnight at 4ºC
Incubation with secondary AB (ab6721 - Goat Anti-Rabbit IgG) 1:2000 for 1h at RT
Developed with ECL for 537s
Results
No nonspecific binding. No background. Clear bands.
Username

Mr. Helder Fonseca

Verified customer

Submitted Oct 07 2014


In the case of ab6721, the storage condition has changed because we removed the glycerol bit from the antibody storage buffer. As a consequence, from now on we recommend to store the antibody at 4 degrees.
For more information on antibody st...

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The dilution factor on the datasheet only a recommendation for starting. The actual dilutions need to be optimized by the scientist since it depends on many factors such expression levels of the target in the samples, primary antibody (clonality, purif...

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The main difference in the 3 secondary antibodies you mentioned are in the applications that they are tested and guaranteed to work in for 6 months as stated in our AbPromise guarantee, the method in which they were purified, and whether or not they ar...

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We are aware of this issue; the brown precipitates as observed are very common with HRP conjugated secondary antibodies so our lab is devising ways to get rid of these precipitates however results are still awaiting.

We and our customers have ...

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I can confirm that for the HRP conjugated antibodies the approach that I recommended is the correct one. Spining the vial down will prevent free HRP to bind to unspecifically to samples and therefore avoid increased background.

If the antibod...

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Please dilute the antibodies in PBS with 1-3% BSA solution.

The rabbit light chain-specific antibody ab99697 should be suitable for your Western blot and I wouldn't expect it to cross-react with the reduced denatured mouse IgG heavy chain, however this antibody has not been tested for reactivity with mouse IgG ...

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The concentration of this antibody is 2 mg/mL, and our vials contain 1 mg of antibody. In this case you would expect 500 uL of antibody in the vial.

We have a ECL Western Blotting Substrate Kit would be compatible with the goat polyclonal secondary antibody ab6721. This comes in two sizes, ab65623 in 50 testes and ab65628 in 500 tests. More information on these products can be found from the follow...

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