Overview

  • Product name
    Mouse on Mouse Polymer IHC Kit
  • Sample type
    Tissue
  • Species reactivity
    Reacts with: Mouse
  • Product overview

    ab127055 is a robust ONE-step polymer system that provides increased sensitivity, time savings and detection simplicity. ab127055 allows the use of mouse antibodies on mouse tissue to obtain excellent signal intensities and clean background. The antibody-antigen interaction is visualized by adding a HRP conjugated secondary antibody followed by a chromogen (e.g. DAB (ab94665)) that yields a permanent brown precipitate. This system is also biotin-free, which eliminates background staining found with traditional biotin-based detection methods and improves signal-to-noise ratios.


    The kit contains a rodent blocking reagent, which is a F(ab) fragment of an anti-mouse / anti-rat secondary antibody. This reagent binds to, and saturates, any endogenous mouse antibodies present in the tissue section. This prevents the binding of the mouse on mouse HRP polyer secondary antibody to endogenous mouse antibodies.

  • Notes

    Additional materials required:

    • Antigen Retrieval Buffer (100X Citrate Buffer pH 6.0) (ab94674)
    • Antigen Retrieval Buffer (100X EDTA Buffer, pH 8.0) (ab94677)
    • Antigen Retrieval Buffer (100X Tris Buffer, pH 10.0) (ab94680)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab94681)
    • DAB substrate (ab94665)
  • Tested applications
    Suitable for: IHC-Pmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab127055 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.

Images

  • Mouse Pan CK (Clone AE1/AE3) antibody staining Pan CK in mouse colon tissue with the use of ab127055.

  • Immunohistochemical analysis with ab127055 showing Dazl and Boll protein expression during mouse fetal ovary development.

    High expression of Dazl (top panel) is seen in germ cells through all stages of development. Boll (bottom panel) is only expressed in germ cells during e15.5.

  • Immunohistochemical analysis of mouse eye using ab127055 to optimize signal to background staining ratio.

     

  • Mouse Pax-5 antibody staining Pax-5 in mouse spleen with the use of the ab127055.

Protocols

References

This product has been referenced in:
  • Nie Y  et al. Palmdelphin promotes myoblast differentiation and muscle regeneration. Sci Rep 7:41608 (2017). Read more (PubMed: 28148961) »
  • Boutilier JK  et al. Gene Expression Networks in the Murine Pulmonary Myocardium Provide Insight into the Pathobiology of Atrial Fibrillation. G3 (Bethesda) 7:2999-3017 (2017). IHC-P ; Mouse . Read more (PubMed: 28720711) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Abreviews
Experimental settings:
• Tissue: cryo sections of decidua from the 11th week of pregnancy.
• Fixation: acetone at room temperature for 5 minutes.
• Blocking: blocking with UV ultra-block (Thermo Fischer Scientific) for 10 min.
• Washing: During IHC protocol after each step, sections were washed three times in TBS-T buffer for 5 min at room temperature.
• Primary antibody: anti neutrophil elastase (Dako), dilution: diluted in antibody diluent (Dako, 1:25) for 30 minutes at room temperature
• Detection Kits (used according to the manufacturer’s instructions):
1. UltraVision LP Detection System from LabVision (detects primary mouse as well as rabbit antibodies)
2. Mouse on mouse from Abcam
3. Mouse on Mouse Kit from Vectorlabs (already 7 years old and still pretty nicely working)
• Development: aminoethylcarbazole (AEC, LabVision) for 4 min at room temperature
• Counterstaining: Mayer’s hemalum
• Mounting: Kaiser’s glycerolgelatine (Merck).
Results:
1. The kit from LabVision worked very well, as we know.
2. The Mouse on Mouse Kit from Abcam did not give any positive signal at all.
3. Also the kit from Vectorlabs, although already 7 years old, gave a pretty nice distinctive signal.

Christopher Trummer

Verified customer

Submitted Sep 10 2018

Mouse on Mouse Polymer IHC kit

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Tissue: paraffin embedded mouse xenograft tumor sections

Antigen retrieval: TE pH9, blocked with rodent block from kit.
Antibody diluent: 2% BSA
Primary antibody: mouse anti-Smooth muscle actin, 1:200, overnight incubation at 4oC
Secondary antibody: Mouse on mouse polymer HRP secondary antibody used at 2x dilution, incubate for 30 minutes at room temperature
Detected with DAB kit from Vector lab

Abcam user community

Verified customer

Submitted Jan 03 2018

Life saving Mouse on Mouse Polymer IHC Kit for IHC

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We have been having an issue getting this elastase staining to work o mouse lung tissue, given that the elastase ab (ab52484) is mouse originated. We've decided to order this kit and give it a try and did not regret ever since. It was a great kit that allows us to clearly visualized histone staining in addition to the other markers. In the following pictogram, elastase+ signal was labeled as red, while the brown cells are neutrophils.

Jim Hsiao

Verified customer

Submitted Dec 19 2017

Postnatal day 6 mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with a 50/50 mixture of Rodent Block and 3% BSA in 0.1% Triton X-100 + PBS. Sections were then incubated for 1h at RT with either (A) no primary antibody or (B) anti-DDX4 (Abcam ab27591) at a 1:100 dilution in a 50/50 mixture of Rodent Block and 3% BSA in 0.1% Triton X-100 + PBS. Sections were then washed 3X with 0.1% Triton X-100 in PBS. Donkey Anti-Mouse 555 (Abcam ab150110) applied at a 1:500 dilution in 3% BSA in 0.1% Triton X-100 + PBS. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.

Mr. Bryan Niedenberger

Verified customer

Submitted Nov 21 2016

Answer

Thank you for your enquiry and your interest in our products.

I understand from your previous e-mail that your customer will perform IHC-P with samples from either rat or rabbit.

I can confirm that ab80436 EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit can be used for primary antibodies raised either in mouse or rabbit. However, if the host species of the primary antibody is the same as the sample species, there is unfortunately a high risk for non-specific background signal.

We have a special kit ab127055 which allows the use of mouse antibodies on mouse tissue to obtain excellent signal intensities and clean background.


I am not sure if I understand what you refer to protocol #7 and protocol #8? Could you please clarify?

If you need any further assistance in the future, please do not hesitate to contact me.

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Question
Answer

Thank you for contacting us.

As requested, I have attached the datasheet for ab7032 as well as our Mouse on Mouse staining protocol. Abcam also carries a Mouse on Mouse staining kit which contains all of the reagents need to perform mouse on mouse staining and has been designed to eliminate any background issues when performing this type of staining. More information about this kit may be found at the following location:

https://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.html

We also have a number of anti-MMP2 antibodies currently on our catalog which are guaranteed in IHC-P on mouse tissues. These products are listed below:

https://www.abcam.com/mmp2-antibody-6e3f8-ab86607.html

https://www.abcam.com/mmp2-antibody-ab110186.html

https://www.abcam.com/mmp2-antibody-ab37150.html



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for contacting us.

This kit will work with mouse IgM antiobodies as well.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for contacting us.

ab127055 kit can be used with any mouse antibody - this mean the kit would be suitable.

Please note ab49536 is tested and guaranteed to be with mouse species however ab57752 is only tested with human samples and as the immunogen is not compatible with mouse species so we cannot guarantee this antibody. Please select a different antibody.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for contacting us.

I have found some information regarding the anti-IL-10 antibodies which we spoke of. To begin, I have found that Mouse IL-10 and Human IL-10 have only a 73% homology which is especially poor in the region around aa13-aa70 of mouse IL-10 (http://www.uniprot.org/uniprot/P18893).

The peptide from which ab9969 was created has 78% identity. Generally we do not expect an antibody to recognize a target if the homology is below 85% but this product is polyclonal and as such may be able to recognize human IL-10 in some lots.



The immunogen used to generate ab33471 is E coli-expressed recombinant mouse IL-10. We have not mapped the epitope of this antibody so we do not know exactly where the antibody will bind. .However this is a monoclonal product and as stated above the homology between IL-1o mouse and human is below the threshold for antibody recognition. As such this may be an appropriate choice but we have not tested in-house for cross reactivity of this antibody andcannot guarantee that it will not recognize human IL-10.



The same statements above can be repeated for ab25073. However as this is a mouse monoclonal and you wish to work in mouse tissue we would highly recommend use of theMouse on Mouse Polymer IHC Kit ab127055.



AB33471 and ab9969 would also qualify for enrollment in our AbTrial testing program. For UNTESTED applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount equal to the value of the product tested. The terms and conditions of this offer can be found here:https://www.abcam.com/AbTrial

I have included links to each of these products below:

AB9969:https://www.abcam.com/il10-antibody-ab9969.html

AB33471:https://www.abcam.com/il10-antibody-jes5-2a5-ab33471.html

AB25073:https://www.abcam.com/il10-antibody-2g101h7-ab25073.html

Mouse on Mouse Kit:https://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.html

I hope this information is helpful to you. If you are interested in testing one of these products in IHC-P please let me know. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Vielen Dank für Ihre Anfrage.

Gerne senden wir Ihnen einen kostenlosen Ersatz. Bitte teilen Sie mir doch mit, wann (ungefähr) Sie den Antikörper gekauft haben, damit ich Ihre Bestellung heraus suchen kann.

Wenn Sie eine "Mouse-on-Mouse" Färbung durchführen möchten, kann es natürlich immer zu einem erhöhten Hintergrund kommen. Ich würde dazu entweder einen Sekundär- AK empfehlen, der nur den Isotyp des Primär- AKerkennt (was den Hintergrund aber nicht komplett verschwinden lässt), oder aber ein speziel fuer diese Zwecke entwickeltes IHC Kit empfehlen:

https://www.abcam.com/index.html?datasheet=127055 (or use the following: https://www.abcam.com/index.html?datasheet=127055).

Ich hoffe, dies hilft Ihnen weiter. Ich freue mich auf Ihre Antwort.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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1-10 of 19 Abreviews or Q&A

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

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