Overview

  • Product name
    Goat Anti-Rabbit IgG H&L (Biotin)
    See all IgG secondary antibodies
  • Host species
    Goat
  • Target species
    Rabbit
  • Specificity
    The antibody used for conjugation reacts with rabbit immunoglobulins of all classes. Cross-reactions as determined by ELISA for the unconjugated antibody (ab182016): Mouse IgG, rat IgG, and chicken IgY, less than 2%. Human IgG, less than 7%.
  • Tested applications
    Suitable for: ELISA, ICC/IF, IP, Flow Cyt, WB, IHC-Fr, IHC-Pmore details
  • Immunogen

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

  • Conjugation
    Biotin

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
    pH: 7.4
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 1% BSA, 30% Glycerol
  • Concentration information loading...
  • Purity
    Affinity purified
  • Purification notes
    Immunogen affinity purified - This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to Biotin.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab207995 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
ELISA 1/20000 - 1/200000.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IHC-P 1/500 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Images

  • IHC image of Histone H4 staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*. Ab207995 Goat Anti-Rabbit IgG H & L (Biotin) was used as the secondary antibody.

    Staining was performed on a Leica BondTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins, before blocking of endogenous biotin using ab64212. The section was then incubated with ab177840, 1/100 dilution, for 15 mins at room temperature, followed by ab207995, 1/2000 dilution, for 15 mins at room temperature. Detection was via an HRP conjugated ABC system and DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ab207995 was tested by direct ELISA, where wells were coated with serially diluted rabbit IgG (1000 – 16 ng/ml) for 2 hours, followed by a 2 hour blocking step (5% BSA). ab207995 (1:20,000 dilution; 2 hours) was added and detected by streptavidin-HRP (ab7403; 1:10,000 dilution; 1 hour). Signal was developed by TMB substrate. Data from duplicates; +/- SD.

  • IHC image of beta Tubulin staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*. Ab207995 Goat Anti-Rabbit IgG H & L (Biotin) was used as the secondary antibody.

    Staining was performed on a Leica BondTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins, before blocking of endogenous biotin using ab64212. The section was then incubated with ab6046, 1/100 dilution, for 15 mins at room temperature, followed by ab207995, 1/1000 dilution, for 15 mins at room temperature. Detection was via an HRP conjugated ABC system and DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Cross-reactivity of the polyclonal secondary antibody ab182016 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. ab182016 was then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT.

    For the batch tested, ab182016 showed a cross-reactivity of 5-7% towards Human IgG and below 2% towards Mouse IgG, Rat IgG and Chicken IgY.

    This data was developed using the unconjugated antibody (ab182016).

  • Cross-reactivity of Goat anti-Rabbit IgG H&L (ab182016) and Goat anti-Rabbit IgG H&L obtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT. This data is from a representative dilution.

    This data was developed using the unconjugated antibody (ab182016).

References

This product has been referenced in:
  • Muraki M & Hirota K Site-specific chemical conjugation of human Fas ligand extracellular domain using trans-cyclooctene - methyltetrazine reactions. BMC Biotechnol 17:56 (2017). Read more (PubMed: 28673349) »
See 1 Publication for this product

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