Mouse on Mouse Polymer IHC Kit (ab269452)
Overview
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Product name
Mouse on Mouse Polymer IHC Kit -
Sample type
Tissue -
Species reactivity
Reacts with: Mouse -
Product overview
Mouse on mouse polymer IHC kit ab269452 allows antibodies raised in mouse to be used on mouse tissues in IHC, without high backgrounds caused by anti-mouse secondary antibody binding to endogenous mouse antibodies in mouse tissues.
The mouse on mouse kit includes an Fab'anti-mouse IgG(H+L) blocking reagent and an HRP polymer conjugated Fab'anti-mouse IgG(H+L) detection reagent.
This kit replaces mouse on mouse kit ab127055. The reagents used in this kit have been carefully optimized to match the performance of ab127055 (see validation images below). This kit does not include hydrogen peroxide blocking reagent: we recommend hydrogen peroxide blocking reagent ab64218.
In the mouse on mouse IHC protocol:
- Cover the tissue section with the blocking reagent (before the addition of the mouse primary antibody) for 30-60 minutes. Wash 2-3 times.
The blocking reagent binds to endogenous mouse antibody in the tissue and prevents the binding of anti-mouse secondary antibody to those sites.
- Incubate with primary antibody of your choice raised in mouse. Wash 2-3 times.
- Incubate with HRP polymer detector reagent for ~15 minutes. Wash 2-3 times.
- Stain with a chromogen (usually DAB, eg. ab64238) for detection.
Otherwise, follow a conventional IHC protocol for DAB staining.
The detector and blocker have <1% reactivity to human and rabbit antibodies.
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Notes
Other products for IHC
Find more kits and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.
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Tested applications
Suitable for: IHC-Pmore details
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 15 ml 60 ml HRP polymer detector (ready to use) 1 x 15ml 1 x 60ml Mouse on mouse blocking reagent (ready to use) 1 x 15ml 1 x 60ml
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab269452 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 |
Use at an assay dependent concentration.
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| Notes |
|---|
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IHC-P
Use at an assay dependent concentration. |
Images
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Mouse on Mouse IHC test with comparison to ab127055
Mouse on mouse polymer IHC kit ab269452 (panel B) was tested using an anti-actin mouse monoclonal on formalin-fixed paraffin embedded mouse tissue sections.
Equivalent staining was seen using the same antibody and serial tissue section with ab127055 (panel C).
A control without primary antibody showed no staining (panel A).
Antigen retrieval was using 1mM EDTA, pH 9.0 for 15 minutes. The detection reagents in the mouse on mouse kits were incubated with the section for 15 minutes.
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Mouse on Mouse IHC test with comparison to ab127055Mouse on mouse polymer IHC kit ab269452 (panel B) was tested using an anti-actin mouse monoclonal on formalin-fixed paraffin embedded mouse tissue sections.
Equivalent staining was seen using the same antibody and serial tissue section with ab127055 (panel C).
A control without primary antibody showed no staining (panel A).
Antigen retrieval was using 1mM EDTA, pH 9.0 for 15 minutes. The detection reagents in the mouse on mouse kits were incubated with the section for 15 minutes.
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Mouse on Mouse IHC test with comparison to ab127055Mouse on mouse polymer IHC kit ab269452 (panel B) was tested using an anti-actin mouse monoclonal on formalin-fixed paraffin embedded mouse tissue sections.
Equivalent staining was seen using the same antibody and serial tissue section with ab127055 (panel C).
A control without primary antibody showed no staining (panel A).
Antigen retrieval was using 1mM EDTA, pH 9.0 for 15 minutes. The detection reagents in the mouse on mouse kits were incubated with the section for 15 minutes.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab269452 has been referenced in 1 publication.
- Kakehashi A et al. Cache Domain Containing 1 Is a Novel Marker of Non-Alcoholic Steatohepatitis-Associated Hepatocarcinogenesis. Cancers (Basel) 13:N/A (2021). PubMed: 33802238