Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab103203 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
ICC/IF Use at an assay dependent concentration.

Target

  • Relevance
    DMP1 is an extracellular matrix protein and a member of the small integrin binding ligand N-linked glycoprotein family. It is critical for proper mineralization of bone and dentin, and is present in diverse cells of bone and tooth tissues. DMP1 contains a large number of acidic domains, multiple phosphorylation sites, a functional arg-gly-asp cell attachment sequence, and a DNA binding domain. DMP1 may also have a dual function during osteoblast differentiation. In the nucleus of undifferentiated osteoblasts the unphosphorylated form acts as a transcriptional component for activation of osteoblast-specific genes like osteocalcin. During the osteoblast to osteocyte transition phase it is phosphorylated and exported into the extracellular matrix, where it regulates nucleation of hydroxyapatite.
  • Cellular localization
    Nucleus. Cytoplasm. Secreted; extracellular space; extracellular matrix. Note=In proliferating preosteoblasts it is nuclear, during early maturation stage is cytoplasmic and in mature osteoblast localizes in the mineralizated matrix. Export from the nucleus of differentiating osteoblast is triggered by the release of calcium from intracellular stores followed by a massive influx of this pool of calcium into the nucleus.
  • Database links
  • Alternative names
    • ARHP antibody
    • ARHR antibody
    • AV020965 antibody
    • Dentin matrix acidic phosphoprotein 1 antibody
    • Dentin matrix protein 1 antibody
    • DENTMAT antibody
    • DMP 1 antibody
    • Dmp antibody
    • MGC130441 antibody
    • PP antibody
    • Serine rich acidic phosphoprotein antibody
    see all

Images

  • Anti-DMP1 antibody (ab103203) at 1 µg/ml + Bone Marrow (Human) Tissue Lysate - adult normal tissue 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.

    Predicted band size: 55 kDa
    Observed band size: 55 kDa
    Additional bands at: 50 kDa (possible truncated form)


    Exposure time: 4 minutes
  • Anti-DMP1 antibody (ab103203) at 1/200 dilution + whole cell lysate prepared from murine bone marrow stromal osteoblasts at 20000 cells

    Secondary
    HRP-conjugated goat polyclonal at 1/120000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 55 kDa
    Observed band size: 55 kDa


    Exposure time: 30 seconds


    Osteoblasts were treated with 50µg/ml L-ascorbic acid, 10 mM betha-glycerophosphate for 7 days.

    See Abreview

References

This product has been referenced in:
  • Gulseren G  et al. Dentin Phosphoprotein Mimetic Peptide Nanofibers Promote Biomineralization. Macromol Biosci N/A:e1800080 (2018). Read more (PubMed: 29745025) »
  • Jiang S  et al. Disruption of kif3a results in defective osteoblastic differentiation in dental mesenchymal stem/precursor cells via the Wnt signaling pathway. Mol Med Rep 14:1891-900 (2016). WB ; Human . Read more (PubMed: 27432616) »
See all 5 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Proteinase K
Sample
Mouse Tissue sections (Tibia)
Specification
Tibia
Permeabilization
Yes - Methanol
Fixative
Paraformaldehyde

Mr. Vasily Gnyubkin

Verified customer

Submitted Sep 11 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (PULP)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
40 µg
Specification
PULP
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Kyungjoo Seong

Verified customer

Submitted Nov 30 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Bone Tissue)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
40 µg
Treatment
100 ug/ml of Insulin
Specification
Bone Tissue
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C

Dr. Francesco Elia Marino

Verified customer

Submitted Oct 16 2015

Question
Answer

Thank you for taking the time to complete our questionnaire. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

In order to better understand the images sent I would appreciate if you could please send the molecular weight markers together with the images. As far as I understand the positive control is showing the correct molecular weight, which is different from your sheep lysates, is that correct?

Since you used this antibody last year in ICC, how was it stored? Was it aliquoted or has it suffered thawing / freezing cycles? Have you used it for ICC again after this time?

Have you tried repeating the experiment using enzymatic inhibitors in your lysis buffer? This may help remove the non specific bands, and if a milder blocking agent is afterwards used, it is possible you may be able to observe the correct bands.

I would also appreciate if you could please provide the order number for this product.

I hope this suggestions help improve the results from the antibody. Please do not hesitate to contact us for further assistance.

Read More

Answer

Thank you for contacting us.

I am sorry to hear that you are experiencing difficulties with this product ab103203 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications / species cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered.

In order to better understand the problem, I would like to send you a questionnaire with some additional questions about the protocol used, as well as the order details. All this information is crucial to help us understand the possible causes of the problem. I would also appreciate if you could please send the Molecular Weight Markers in the image.

I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question
Answer

Thank you for contacting us.
We have many antibodies that specifically stain endothelial and osteoblast marker proteins. The most popular antibodies are as follows;
Osteocalcin, DMP-1 and Osteopontin are Osteoblast marker proteins, the antibodies against these are;
ab13420, ab13418, ab13421, ab14173
ab103203, ab81985, ab76632 and 82351
ab8448, ab69498, ab91655 and ab63856
anti Von antibodies are;
ab6994, ab11713 and ab9378 are the best antibodies available.
Please click on the link to explore secondary antibodies with different conjugates;

https://www.abcam.com/index.html?pageconfig=productmap&cl=918
As per your email I think the application you are interested in, is double immunostaining. Please find attached the general protocols for double immunostaining.
https://www.abcam.com/index.html?pageconfig=resource&rid=11458
https://www.abcam.com/index.html?pageconfig=resource&rid=11459
Double immunostaining can be divided in two types:
Labelling using fluorochromes: The commonest fluorochrome available is FITC which gives green colour, TRITC and Texas red which gives red color. Other fluorochromes are Dylight 488 (Green), Dylight 633 (red) and so on. Please click the for complete list of fluorochromes (https://docs.abcam.com/pdf/immunology/fluorochromes_table_updated.pdf). The secondary antibodies conjugated to these fluorochromes can be used to detect 2 marker proteins same time in double immunolabelling. Please be advised that while selecting the fluorophores the absorption maxima of one dye should not overlap with other.
Labelling using enzymatic methods: Enzyme labelled secondary antibodies are used in this method. The primary antibodies should either be from different species (e.g. rabbit or mouse) or both from same species but different in isotypes. When using secondary antibodies please use one HRP conjugated and other AP (alkaline phosphatase) conjugated ab so using substrate for each enzyme will give two colour staining.
Things to remember in double staining
- The primary antibodies should have different host
- If the host is same they should be different isotypes e.g. IgG1a and IgG1 or IgG1 and IgG3 etc.
- If the two targets are expressed on same part of cell then fluorochrome is best method, if the markers are expressed on different part of cells e.g. membrane or nucleus then enzymatic method is best.
- Use the optimized dilution of primary antibodies.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Sheep Cell (Ovine fibroblasts from skin biopsies)
Specification
Ovine fibroblasts from skin biopsies
Fixative
Paraformaldehyde
Permeabilization
No

Abcam user community

Verified customer

Submitted May 19 2011

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Bone marrow stromal cells/osteoblasts)
Loading amount
20000 cells
Specification
Bone marrow stromal cells/osteoblasts
Treatment
50microg/ml L-ascorbic acid, 10 mM betha-glycerophosphate for 7days
Gel Running Conditions
Reduced Denaturing (4-10%)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Mar 23 2011

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

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