Overview

  • Product name

    Anti-Fibromodulin antibody
    See all Fibromodulin primary antibodies
  • Description

    Rabbit polyclonal to Fibromodulin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Fr, IHC-P, ELISAmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit, Horse, Guinea pig, Cow, Cat, Dog
  • Immunogen

    Synthetic peptide corresponding to a region within internal amino acids 108-157 (

    KYVYFQNNQITSIQEGVFDNATGLLWIALHGNQITSDKVGRKVFSKLRHL

    ) of human Fibromodulin (NP_002014).

  • Positive control

    • 721_B cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab81443 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 43 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-Fr Use at an assay dependent concentration. PubMed: 21939397
IHC-P 1/200.
ELISA Use at an assay dependent concentration.

Target

Images

  • ab81443 staining Fibromodulin in Mouse aorta tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% Formalin and blocked with 10% serum for 30 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200 in 10% Normal Goat serum) for 16 hours at 4°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/375) was used as the secondary antibody.

    See Abreview

  • Anti-Fibromodulin antibody (ab81443) at 1 µg/ml + 721_B cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 43 kDa
    Observed band size: 43 kDa



    Gel concentration: 12%.
  • ICC/IF image of ab81443 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab81443, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Anti-Fibromodulin antibody (ab81443) at 1/500 dilution + Equine Cartilage Explants

    Predicted band size: 43 kDa



    WB Suggested Anti-SFN antibody (ab87209) Titration: 2 µg/ml Positive Control: sea urchin embryo

References

This product has been referenced in:

  • Li CJ  et al. Long noncoding RNA Bmncr regulates mesenchymal stem cell fate during skeletal aging. J Clin Invest 128:5251-5266 (2018). Read more (PubMed: 30352426) »
  • Dyment NA  et al. The paratenon contributes to scleraxis-expressing cells during patellar tendon healing. PLoS One 8:e59944 (2013). IHC-Fr ; Mouse . Read more (PubMed: 23555841) »
See all 5 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Heart)
Specification
Heart

Abcam user community

Verified customer

Submitted Apr 18 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Chicken Tissue sections (HH31 wild type chick embryo trunk -sagittal sectio)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
HH31 wild type chick embryo trunk -sagittal sectio
Blocking step
BSA + 5% heat-inactivated goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1000µg/mL · Temperature: 20-25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 14 2015

Answer

Thank you for that information.
As discussed, I can arrange for the ab81443 of a different lot number to be sent to you. However I will just need to check your order details for the original vial of ab81443. I have the following information, does this match your order and is this the delivery address you would like the antibody sent to?
Order number xxxx Purchase order number xxxxxx
Delivery address:
xxxxxxx
Once I have this confirmation I will arrange for the shipment. I look forward to receiving your reply.

Read More

Answer

Thank you for your reply.
This has made understanding the problems you have encountered and what may be contributing to them easier to understand. I cannot be sure what is causing this problem, but from what you have described, I would suggest it may either be the fixation or the antigen retrieval currently used.
If the tissue is over fixed, this can leads to the antibody not being able to recognise the protein. Alternatively, non-optimal antigen retrieval can also lead to the epitope not being exposed for detection. Therefore, can i ask the following:
1. How was the tissue harvested and fixed?
2. When optimising the antigen retrieval, which methods were used? You have mentioned the buffers but the method of heating (microwave, pressure cooker) and for what times were the incubations tried?
I cannot rule out either that the antibody itself has become inactivated for some reason. I understand that you have used a lot of time and effort in trying to optimise the results and I would have expected there to be some staining from the information you have shared with me. I am sorry to for the inconvenience this problem is causing you.
If you would like, I can send you a new vial from a different lot of ab81443 free of charge to try? Alternatively, I can send you an alternative antibody altogether. Unfortunately, we do not currently have any other anti-Fibromodulin antibodies available which have been shown to work in IHC but any other antibody from our catalogue. Or I can understand if you would prefer a refund or credit note to be issue.
I look forward to receiving your reply.

Read More

Answer

Thank you for contacting us and raising your concerns in regards to the anti-Fibromodulin antibody (ab81443).
I am sorry to hear you are experiencing difficulties with this product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
It seems from the details you have provided that the antibody should have worked with your samples. In order to try and understand more fully why this has not been the case, would you mind providing me with a few more details of the experiment being carried out? To this end I have attached a questionnaire.
Also, you mentioned you have been using another anti-Fibromodulin antibody with better results. Could you let me know which antibody this is (supplier and catalogue number)?
Once I have received the completed questionnaire, I will look at the protocol and see if there are any suggestions I can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary.
If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.
I look forward to receiving your reply.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (aorta)
Specification
aorta
Fixative
10% formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Aug 03 2012

Question
Answer

DISCOUNT CODE: *
Expiration date: *
I am very pleased to hear you would like to accept our offer and test ab81443 in IHC-P. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for IHC-P and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.
Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.
The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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