Anti-Collagen X antibody (ab58632)

Rabbit polyclonal Collagen X antibody. Validated in WB, IP, IHC, ICC/IF and tested in Mouse, Rat, Rabbit, Cow, Human. Cited in 96 publication(s). Independently reviewed in 9 review(s).

Overview

  • Product name

  • Description

    Rabbit polyclonal to Collagen X
  • Host species

    Rabbit
  • Specificity

    ab58632 recognizes type X collagen. Exhibits slight cross-reactivity with fibronectin and type II and type IX collagen. Does not cross-react with type I, type III, or type XI collagen.
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-Frmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Rabbit, Cow, Human
  • Immunogen

    Full length native protein (purified) Type X collagen from rat chondrosarcoma cells

Properties

Applications

Our Abpromise guarantee covers the use of ab58632 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 at an assay dependent concentration.
WB 1/100 - 1/300.
IP 1/20 - 1/40.
IHC-Fr Use at an assay dependent concentration. Enzymatic or heat (41°C) pretreatment is recommended.

Target

  • Function

    Type X collagen is a product of hyperthrophic chondrotocytes and has been localized to presumptive mineralization zones of hyaline cartilage.
  • Involvement in disease

    Defects in COL10A1 are the cause of Schmid type metaphyseal chondrodysplasia (SMCD) [MIM:156500]. SMCD is a dominantly inherited disorder of the osseous skeleton. The cardinal features of the phenotype are mild short stature, coxa vara and a waddling gait. Radiography usually shows sclerosis of the ribs, flaring of the metaphyses, and a wide irregular growth plate, especially of the knees. A variant form of SMCD is spondylometaphyseal dysplasia Japanese type. It is characterized by spinal involvement comprising mild platyspondyly, vertebral body abnormalities, and end-plate irregularity.
  • Sequence similarities

    Contains 1 C1q domain.
  • Post-translational
    modifications

    Prolines at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Database links

  • Alternative names

    • COAA1_HUMAN antibody
    • Col10a 1 antibody
    • COL10A1 antibody
    • Collagen alpha 1(X) chain antibody
    • Collagen alpha-1(X) chain antibody
    • Collagen type X alpha 1 (Schmid metaphyseal chondrodysplasia) antibody
    • Collagen type X alpha 1 antibody
    • Collagen X alpha 1 polypeptide antibody
    • CollagenX antibody
    • fa66d11 antibody
    • fb10c08 antibody
    • OTTHUMP00000040411 antibody
    • Procollagen type X alpha 1 antibody
    • Schmid metaphyseal chondrodysplasia antibody
    • wu:fa66d11 antibody
    • wu:fb10c08 antibody
    see all

Images

  • Immunofluorescence analysis of mouse bone marrow stromal cells, staining Collagen X with ab58632.

    Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 before incubating with primary antibody (1/50) overnight. An AlexaFluor®488-conjugated anti-rabbit IgG (1/500) was used as the secondary antibody.
  • Anti-Collagen X antibody (ab58632) at 1/500 dilution + HT 1080 (Human fibrosarcoma) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 66 + 64 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 37 kDa, 50 kDa, 98 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 150 seconds


    The band observed at 64 kDa could potentially be a cleaved form of Collagen X due to the presence of a 18 amino acid signal peptide.

References

This product has been referenced in:

  • Liao L  et al. Deletion of Runx2 in condylar chondrocytes disrupts TMJ tissue homeostasis. J Cell Physiol 234:3436-3444 (2019). Read more (PubMed: 30387127) »
  • Sun AR  et al. Pro-resolving lipid mediator ameliorates obesity induced osteoarthritis by regulating synovial macrophage polarisation. Sci Rep 9:426 (2019). Read more (PubMed: 30674985) »
See all 111 Publications for this product

Customer reviews and Q&As

1-9 of 9 Q&A

Answer

Antibody ab58632 is our recommendation, given the number of references that have used it for IHC of FFPE tissue. Here is a link to the list of references, annotated "IHC-P" for paraffin.

https://www.abcam.com/collagen-x-antibody-ab58632-references.html

The four other antibodies that have been tested in this application should be effective, at least for the species that are listed on the datasheets, but they do not have as much data. My second choice would be the mouse monoclonal ab49945, depending on the species of your samples. For instance, I would not recommend it for staining mouse tissue, if there is endogenous mouse IgG in the sample that a secondary anti-mouse IgG antibody might react with. However, two references do have data from immuno-compromised mice.

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Question
Answer

Thank you for contacting us. The anti-Collagen X antibody ab58632 is a whole serum antibody and the concentration has not been determined. Generally concentrations are only relevant for purified antibodies, since products from ascites or whole serum may contains proteins other than the target protein. We recommend that you follow the suggested dilution for your application, which for IHC is 1/50-1/100.

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

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab49945with the order number 1115958. You can expect to receive this on July 2nd, but to check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

Thank you for contacting us and for your interest in our products.

I am sorry to confirm that as far as we are aware, ab58632 has never been tested with samples from sheep. All tested and guaranteedspecies cross-reactivity information is stated on our datasheets, and these are updated as soon as any new information is brought to our attention.

If you would like to test the antibody in this untested species, please contact our Scientific Support team by replying to this message prior to the purchase as you may be eligible for our testing discount program.

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. 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 me for any further advice or information in this regard.

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Answer

Thank you for contacting us.

The antibody is whole, unpurified antiserum, and the antibody concentration is not determined. Dilutions for some common applications are suggested on the datasheet. The appropriate isotype control for this antibody is whole normal rabbit serum, such as ab7487, diluted by the same factor as ab58632.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=7487).

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

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Answer

Thank you for your inquiry. Please find a list of possible positive controls below: ab2426: skin, lymph nodes Click here (or use the following: https://www.abcam.com/index.html?datasheet=2426). ab3773: cartilage samples Click here (or use the following: https://www.abcam.com/index.html?datasheet=3773). ab58632: Type X collagen is a product of hyperthrophic chondrotocytes and has been localized to presumptive mineralization zones of hyaline cartilage. Click here (or use the following: https://www.abcam.com/index.html?datasheet=58632). ab66025: The recommended positive control tissues are rat head tissue, rat embryo tissue, human osteosarcoma tissue, human glioma tissue, human intestinal cancer tissue. Click here (or use the following: https://www.abcam.com/index.html?datasheet=66025). ab102711: Specifically expressed in osteoblasts Click here (or use the following: https://www.abcam.com/index.html?datasheet=102711). I hope this information is helpful for your customer. I wish them good luck with their experiments.

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Answer

Thank you for your call yesterday and for your patience while I have been in touch with the lab. For ab58632, we recommend using the antibody at 1:50-1:100 and incubating overnight. The sections were treated with 0.25% trypsin in 1 mM EDTA for 3 minutes at 37C to expose the epitope. These alterations should improve the staining results with the antibody, however if the problem persists, I would be happy to send a different antibody to try or alternatively issue a credit or refund. Please keep me updated about any new results, and I look forward to hearing from you.

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Answer

Thank you for your response. It is always worth running a "no primary" (only secondary antibody) control to see/check if the secondary antibody alone may cause non-specific signal. In order to reduce the any potential non-specific signal, I would advise the following steps: 1) block the non-specific binding sites with 5% BSA (in TBS-T) for or with 10 % goat serum (host species of the secondary antibody) 1 hr at RT, 2) block endogenous peroxidase activity with H2O2 , You can find some more information on blocking at this site: https://www.abcam.com/index.html?pageconfig=resource&rid=11494 I hope this helps and if I can assist further, please do not hesitate to contact me.

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Answer

Thank you for your enquiry and your interest. I can confirm that ab58632 has been tested and characterized for IHC (both on frozen and on formalin-embedded tissue sections) and it recognizes Mouse, Rat, Rabbit, Cow, Human Collagen X. Since the isotype is rabbit IgG, the secondary antibody should recognize this type of immunoglobulin. Using our advanced search engine, you can conduct a search for the most suitable secondary antibodies i.e. select the host species, conjugation, purification etc. https://www.abcam.com/index.html?pageconfig=productmap&cl=918 If you need any further assistance in the future, please do not hesitate to contact me.

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