Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866)

Overview

  • Product nameAnti-Rabbit IgG VHH Single Domain Antibody (HRP)
    See all Rabbit IgG secondary antibodies
  • Target speciesRabbit
  • Tested applicationsSuitable for: WB, IHC-P, ELISAmore details
  • Immunogen

    Other Immunogen Type corresponding to Rabbit IgG. Immunogen was Fc region of Rabbit IgG

  • ConjugationHRP

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at 4°C (stable for up to 12 months). Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage bufferpH: 7.4
    Preservative: 0.1% Proclin
    Constituents: PBS, Sodium chloride, Sodium phosphate, 30% Glycerol, 1% BSA
  • Concentration information loading...
  • PurityPurified via His tag
  • Purification notesThis product is a recombinant protein produced in E. coli.
  • ClonalityMonoclonal
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab191866 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
WB Use at an assay dependent concentration.
IHC-P Use a concentration of 0.02 - 0.2 µg/ml.
ELISA Use at an assay dependent concentration.

Anti-Rabbit IgG VHH Single Domain Antibody (HRP) images

  • IHC image of Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) staining in formalin fixed paraffin embedded normal human colon tissue section.

    The section was dewaxed and then pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with rabbit polyclonal antibody to beta tubulin (ab6046, 0.5µg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. Endogenous peroxidases were quenched using 1.6% (v/v) hydrogen peroxide in TBS containing 0.025% (v/v) Triton X-100 for 30 minutes at room temperature, with agitation. The secondary antibody, Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866, 0.125µg/ml) was then applied for 1 hour at room temperature in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA before being developed for 10 minutes at room temperature using Steady DAB/Plus (ab103723). The section was then counterstained with hematoxylin and mounted with DPX.

    The negative control (secondary antibody only, no primary) inset shows no staining, demonstrating secondary antibody specificity.

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

  • All lanes : Anti-NRG1 type III antibody (ab23248) at 1 µg/ml

    Lane 1 : Mouse Brain Tissue Lysate
    Lane 2 : Rat Brain Tissue Lysate
    Lane 3 : Mouse Brain Tissue Lysate
    Lane 4 : Rat Brain Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1 - 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml
    Lanes 3 - 4 : Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) at 0.05 µg/ml

    Developed using the ECL technique

    Performed under reducing conditions.
  • IHC image of Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) staining in formalin fixed paraffin embedded normal human cerebellum tissue section.

    The section was dewaxed and then pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with rabbit monoclonal antibody [EPR12763] to NeuN (ab177487, 0.1µg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. Endogenous peroxidases were quenched using 1.6% (v/v) hydrogen peroxide in TBS containing 0.025% (v/v) Triton X-100 for 30 minutes at room temperature, with agitation. The secondary antibody, Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866, 1.0µg/ml) was then applied for 1 hour at room temperature in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA before being developed for 10 minutes at room temperature using Steady DAB/Plus (ab103723). The section was then counterstained with hematoxylin and mounted with DPX.

    The negative control (secondary antibody only, no primary) inset shows no staining, demonstrating secondary antibody specificity.

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

  • Top left: IHC image of Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) staining in formalin fixed paraffin embedded normal human colon tissue section.

    The section was dewaxed and then pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with rabbit polyclonal antibody to Ki67 (ab15580, 0.1µg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. Endogenous peroxidases were quenched using 1.6% (v/v) hydrogen peroxide in TBS containing 0.025% (v/v) Triton X-100 for 30 minutes at room temperature, with agitation. The secondary antibody, Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866, 0.1µg/ml) was then applied for 1 hour at room temperature in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA before being developed for 10 minutes at room temperature using Steady DAB/Plus (ab103723). The section was then counterstained with hematoxylin and mounted with DPX.

    Top right: this image shares the same experimental design parameters, except the secondary antibody was ab97051, goat anti-rabbit IgG H&L (HRP) (0.1µg/ml). This demonstrates the improved definition of staining given by VHH Single Domain Antibodies over conventional secondaries.

    Bottom right: this image shares the same experimental design parameters but is a negative control (no primary antibody) for ab97051, demonstrating specificity of the goat anti-rabbit secondary antibody.

    Bottom left: this image shares the same experimental design parameters but is a negative control (no primary antibody) for ab191866, demonstrating the specificity of the VHH-Single Domain Antibody.

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

  • IHC image of Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) staining in formalin fixed paraffin embedded normal human colon tissue section.

    The section was dewaxed and then pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with rabbit polyclonal antibody to Ki67 (ab15580, 0.1µg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. Endogenous peroxidases were quenched using 1.6% (v/v) hydrogen peroxide in TBS containing 0.025% (v/v) Triton X-100 for 30 minutes at room temperature, with agitation. The secondary antibody, Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866, 0.025µg/ml) was then applied for 1 hour at room temperature in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA before being developed for 10 minutes at room temperature using Steady DAB/Plus (ab103723). The section was then counterstained with hematoxylin and mounted with DPX.

    The negative control (secondary antibody only, no primary) inset shows no staining, demonstrating secondary antibody specificity.

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

  • IHC image of Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) staining in formalin fixed paraffin embedded normal human colon tissue section.

    The section was dewaxed and then pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with rabbit polyclonal antibody to VDAC1 (ab15895, 1/1000 dilution) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. Endogenous peroxidases were quenched using 1.6% (v/v) hydrogen peroxide in TBS containing 0.025% (v/v) Triton X-100 for 30 minutes at room temperature, with agitation. The secondary antibody, Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866, 0.125µg/ml) was then applied for 1 hour at room temperature in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA before being developed for 10 minutes at room temperature using Steady DAB/Plus (ab103723). The section was then counterstained with hematoxylin and mounted with DPX.

    The negative control (secondary antibody only, no primary) insert shows no staining, demonstrating secondary antibody specificity.

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

  • All lanes : Anti-GAPDH antibody (ab37168) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : Jurkat Whole Cell Lysate
    Lane 3 : HeLa Whole Cell Lysate
    Lane 4 : Jurkat Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1 - 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.02 µg/ml
    Lanes 3 - 4 : Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) at 0.02 µg/ml

    Developed using the ECL technique

    Performed under reducing conditions.

References for Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866)

This product has been referenced in:
  • Pang X & Hou X Synergistic protective effect of FTY720 and vitamin E against simulated cerebral ischemia in vitro. Mol Med Rep N/A:N/A (2017). Read more (PubMed: 28498446) »
  • Chen WQ  et al. Influences of PON1 on Airway Inflammation and Remodeling in Bronchial Asthma. J Cell Biochem N/A:N/A (2017). Read more (PubMed: 28657647) »

See all 18 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
The mouse sciatic nerve paraffin sections was dewaxed and rehydrated. Prior to IHC detection, the sections was pre-treated by steaming in 10 mM citrate buffer (pH6.0) for 20 min. Protein non-specific binding was blocked by incubated in 10% goat sera plus 2xcasin for 30min. The sections were then incubated with rabbit anti-beta tubulin III (ab18207) for 1.5 hr at room temp (at 1:1,000 in PBS plus 1xcasein). After washing in PBST (0.05% Tween 20) two times, 5 min each, the sections were incubated with the 2nd antibody, anti-rabbit IgG VHH Single Domain_HRP (ab191866) for 30 min at room temp. After washing in PBST two times, 5 min each, the sections were proceeded with color development with Nova Red as chromogen and then counterstained briefly with hematoxylin and mounted with Permount.
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Submitted Sep 08 2015

Application Western blot
Experimental conditions:
MCF-7 and MDA-MB-231 whole cell lysates extracted with RIPA buffer.
10% SDS-PAGE, 50,000 cells loaded into wells.
Blocking 1h with 5% milk in TBS at RT.
Incubation with primary anti-Vimentin (ab92547) 1:5000 overnight at 4ºC.
Incubation with secondary antibody 0.05ug/ml for 1h at RT.
Developed with ECL for 5 minutes.

Results:
No nonspecific binding. No background. Clear bands.
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Submitted Nov 19 2014

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