概述

  • 产品名称

    小鼠on小鼠Polymer IHC试剂盒
    参阅全部 Mouse on Mouse IHC 试剂盒
  • 样品类型

    Tissue
  • 种属反应性

    与反应: Mouse
  • 产品概述

    This product is being replaced by Mouse on mouse polymer IHC kit ab269452, only a few remaining units of stock of this product are available. Please then buy ab269452 as an alternative; it has been carefully tested for equivalent performance. ab269452 does not contain hydrogen peroxide blocking reagent, we recommend hydrogen peroxide blocking reagent ab64218.


     


    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 (ab64238)) 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 mouse blocking reagent, which is a F(ab) fragment of an anti-mouse 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.

  • 说明

    Additional materials required:

    • Antigen Retrieval Buffer (100X Citrate Buffer pH 6.0) (ab93678)
    • Antigen Retrieval Buffer (100X EDTA Buffer, pH 8.0) (ab93680)
    • Antigen Retrieval Buffer (100X Tris Buffer, pH 10.0) (ab93682)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
    • DAB substrate (ab64238)


    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.

  • 经测试应用

    适用于: IHC-Pmore details

性能

应用

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.

应用 Ab评论 说明
IHC-P Use at an assay dependent concentration.

图片

  • 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.

实验方案

文献

This product has been referenced in:

  • Annese T  et al. a-Methyl-prednisolone normalizes the PKC mediated brain angiogenesis in dystrophic mdx mice. Brain Res Bull 147:69-77 (2019). Read more (PubMed: 30711624) »
  • Taylor-Whiteley TR  et al. Recapitulating Parkinson's disease pathology in a three-dimensional human neural cell culture model. Dis Model Mech 12:N/A (2019). Read more (PubMed: 30926586) »
See all 13 Publications for this product

客户评价及客户问答

11-20 of 20 Abreviews or Q&A

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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Answer

Thank you for contacting us.

You had previously been in contact with my colleague heather regarding staining observed in your negative control using these mouse monoclonal Cytokeratin antibodies on mouse tissue. To clarify, is your negative control from a tissue that is not expected to express cytokeratin or is it an isotype control or a no primary antibody control?

When using an anti-mouse secondary antibody on mouse tissue, background staining is often observed due to cross-reactivity with endogenous mouse IgGs in the sample. This can be checked by testing a no-primary antibody control slide.

If you are observing mouse on mouse background staining, this can be avoided by either using a mouse on mouse (MOM) staining kit, such as ab127055. The background can also be blocked by adding an incubation with a F(ab) fragment unconjugated anti-mouse secondary antibody, such as ab6668. The F(ab) fragment will coat any endogenous IgGs in your sample, and can be done just before the serum block at a 1:500 dilution for 1 hour at room temperature.

I hope this helps, if not, please let me know and I will be happy to help you further.

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Answer

Thank you for contacting Abcam.

I have searched our catalogue, but cannot find any markers specific to MC-38 cells. Do you of any of any specific proteins that are only expressed in these types of cells? As knowing this, would help us find a suitable antibody for your needs.

I also performed a literature search and found the following article that you may find helpful:

http://www.jci.org/articles/view/36046



If there is anything else I can help you with, or if you determine a target protein that you would like to detect, please let me know and I will be happy to do a further search for you.

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Answer

Thank you for contacting us.

I have checked the SwissProt page for the OX-2 membrane glycoprotein, and it looks it is a ligand for CD200R1.Therefore, ab104701 is not the antibody I would like to recommend for your experiments.

However, we have a matching antibody in our catalogue:ab34097 Anti-CD200R antibody [MRC OX102]

https://www.abcam.com/index.html?datasheet=34097 (or use the following: https://www.abcam.com/index.html?datasheet=34097).

Checking the datasheet, I noticed thatab34097 hasn't been tested on mouse.Therefore, I can offer a discount off a future purchase if you buy ab34097 now, test it in mouse and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab34097 in mouse. I will then send a discount code. This code must be issued before purchasing ab34097 so please wait for my reply before ordering.

2. Purchase ab34097 either by phone, fax, or online (www.abcam.com).

3. Test it in mouse.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab34097 turns out to be unsuitable for mouse, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

I know that you wanted a polyclonal antibody from another species, but you could solve this problem by incubating the two primaries + their secondaries after each other.

In addition, we would be delighted if you could write an Abreview about the ab93315 Anti-CD200 / OX2 antibody [OX-2]as well. Unfortunately, as the antibody already has been ordered we cannot issue a testing discount in this case I am afraid.


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

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

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Answer

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

In addition, we have a Polymer IHC Kit which tackles exactly the problem of a high background which arises from this kind of staining (mouse-on-mouse staining):

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

ab127055 allows the use of mouse antibodies on mouse tissue to obtain excellent signal intensities and clean background.

Another alternative i could recommend would be using three antibodies in total: Use a Fab-fragment anti-mouse antibody against the H+L chain to block the endogenous mouse IgGs, than incubate with your primaries and (as you suggested) use an isotype specific secondary antibody.

A Fab fragment antibody has the advantage that it can "cover" the endogenous mouse IgG tighter than a whole IgG antibody.

https://www.abcam.com/index.html?datasheet=6668 (or use the following: https://www.abcam.com/index.html?datasheet=6668).



I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Answer

Thank you for your enquiry.

According to the developer of the kit, the mouse-on-mouse IHC kit will be effective with IgM primary antibodies. The anti-Spamolytic Polypeptide antibody ab49536 is IgM isotype.

Please contact us if you have any other questions or concerns.

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Answer

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Answer

Thank you for contacting us.

Indeed, a mouse-on mouse staining can be quite tricky; however, it is not impossible. The background will depend on the fixation method ( perfusion fixed tissue vs. normally fixed tissue), on the tissue itself ( eg. kidneys vs. bone) and the experimental settings.


One way to make sure to decrease or to completelyerase the background is to block the endogenous mouse IgG with a non- conjugated secondary antibody before you incubate with your target antibody. I would suggest to use a Fab or F(ab)2 fragment antibody in this case as they will bound in a higher density than whole IgG antibodies.

You can find the aforementioned antibodies here:


https://www.abcam.com/index.html?datasheet=98754 (or use the following: https://www.abcam.com/index.html?datasheet=98754).

https://www.abcam.com/index.html?datasheet=6668 (or use the following: https://www.abcam.com/index.html?datasheet=6668).



Another way is to use our hassle- free Mouse on Mouse (M.O.M) Polymer IHC Kit, which contains a special rodent block solution:

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


In addition, we also received an Abreview (17 July 2009) from one of our customers who rated this antibody highly in IHC. You can contact the author via our website if you like.



I hope this information is helpful to you. 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

Thank you for your enquiry and your interest.
Recently, we have launched a new kit (ab127055) Mouse on Mouse (M.O.M) Polymer IHC Kit which allows the use of mouse antibodies on mouse tissue to obtain excellent signal intensities and clean background. It is a ONE-step polymer system that provides increased sensitivity, time savings and detection simplicity.
You may be interested in this new reagent:
https://www.abcam.com/mouse-on-mouse-mom-polymer-ihc-kit-ab127055.html
I hope this helps and if I can assist further, please do not hesitate to contact me.

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Question

Thank you for your quick response. Yes , we tried the staining as per recommended protocol and conditions/ dilutions provided by the abcam with both the antibodies with frozen mouse tissue.



The nature of the problem is that the staining with Twist Ab looks similar and non specific as it appears with the isotype control mouse antibody.Same is the case with the C4d ab that there looks no difference in staining with C4D Ab and with comparable isotype control.





Rehan Kahloon MD
Division of Pulmonary Allergy and Critical Care
Department of Medicine
University of Pittsburgh Medical Center
kahloonra@upmc.edu
412-716-9311


________________________________
From: technical@abcam.com [technical@abcam.com]
Sent: Thursday, April 19, 2012 5:12 PM
To: Kahloon, Rehan
Subject: Reply from Abcam to your enquiry regarding ab36075, ab50887 [CCE3700093]

[abcam: discover more]

Dear Rehan

Thank you for contacting Abcam.

Under our Abpromise we do guarantee our products to work as stated on the datasheet and if for whatever reason they do not work as stated and we cannot help resolve the issue, then we will be happy to either replace or refund the cost of the product. The Abpromise is good for 6 months.

To be able to better assist you in this matter, could you please provide me with information as to how you were using these antibodies, eg western blot, IHC-P, IP etc and also which species your sample came from. If you could also include a brief summary of the nature of the problems you are having, as well as the protocol that you have been using, that would be most helpful.

I look forward to your reply and helping you resolve these issues.

Help us improve our service.
Rate your experience with us today.

Best regards,
John

John Constable
Scientific Support Specialist
Abcam Inc.
www.abcam.com

Your original inquiry to Abcam:

Dear Customer Service,



I am writing in reference to our experience with two of the Antibodies we recently ordered from Abcam.



1- Anti- Twist Ab Catalog 50887 P.O. number 1139131.

2-C4D Antibody Ab Catalog Number ab 36075 P.O. Number 1135999.



Despite repeated attempts and following all the optimal protocols provided, we have not be able to make these antibodies work .



It would be great if we can get our credit back or have them returned, since we believe they are not effective.



Thanks







Rehan Kahloon MD
Division of Pulmonary Allergy and Critical Care
Department of Medicine
University of Pittsburgh Medical Center
WBST1200-23B
200 Lothrop Street.
Pittsburgh,PA.
kahloonra@upmc.edu
412-716-9311







Abcam Customer Services and Scientific Support Team
www.abcam.com/technical

[CCE3700093]




Discover more at abcam.com

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Answer

Thank you for your response.

I am sorry to hear that you have been experiencing problems when using our products. Our anti-Twist antibody in guaranteed to work in mouse tissues in IHC-Fr. We have added recommended dilutions or specific protocols for this producthowever there are customer’s reviews in our Abreview section which do give some protocol information that these customers have used. I noticed differences between these Abreview protocols and was wondering if you could provide me with the protocol steps which you have used, any troubleshooting step that you taken and any images should you have these. If we cannot solve the issues which you are experiencing, I will be glad to send you a free of charge replacement, credit note or refund under our Abpromise guarantee.

We currently do not recommend using ab36075 anti-C4d on mouse tissues. The homology for the immunogen used to create this product and the mouse protein is below the threshold for cross reactivity. As such, I am very happy to look over your protocol and provide tips and suggestions which may help. However this product would not be covered by our Abpromise.

You are doing mouse on mouse staining with these products.Staining of mouse tissue using mouse antibody is a complicated process as high levels of background are often observed. It is notoriously difficult to eliminate this background.I would strongly recommend these tips for eliminating background when staining mouse tissue with a mouse monoclonal.

-1. Block endogenous IgG withunconjugated Fab fragment Anti-Mouse IgG (H+L) for 1 hr at room temperature, or overnight 4°C

-2. Block endogenous Fc receptors withF (ab) monomeric secondary antibodies

-3. Incubate sections with 1% Triton (in PBS) at room temperature for 30 min to ‘clean’ the tissue.

-4. Use TBS –Tween 20 as a washing buffer. This often gives less background than using PBS-Tween 20.

Further information may be found at:https://www.abcam.com/index.html?pageconfig=resource&rid=11465



Alternately I would suggest using theMouse on Mouse (M.O.M) Polymer IHC Kit, ab127055 which is designed toallow the use ofmouse antibodies on mouse tissueto obtain excellent signal intensities and clean background.(https://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.html).





I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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11-20 of 20 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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