Overview

  • Product name
  • Description
    Goat polyclonal to Asporin
  • Host species
    Goat
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide:

    C-IHENKVKKIQKDT

    , corresponding to Internal sequence amino acids 182-194 of Human Asporin. (Peptide available as ab134013.)

  • Positive control
    • Human bone marrow lysate.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 0.5% BSA, Tris buffered saline, pH 7.3
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab31303 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 - 3 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 43 kDa).Can be blocked with Human Asporin peptide (ab134013).
IHC-P Use at an assay dependent dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. PubMed: 20541802
ICC/IF Use at an assay dependent dilution.
ELISA Use at an assay dependent concentration.

Target

  • Function
    Negatively regulates periodontal ligament (PDL) differentiation and mineralization to ensure that the PDL is not ossified and to maintain homeostasis of the tooth-supporting system. Inhibits BMP2-induced cytodifferentiation of PDL cells by preventing its binding to BMPR1B/BMP type-1B receptor, resulting in inhibition of BMP-dependent activation of SMAD proteins (By similarity). Critical regulator of TGF-beta in articular cartilage and plays an essential role in cartilage homeostasis and osteoarthritis (OA) pathogenesis. Negatively regulates chondrogenesis in the articular cartilage by blocking the TGF-beta/receptor interaction on the cell surface and inhibiting the canonical TGF-beta/Smad signal. Binds calcium and plays a role in osteoblast-driven collagen biomineralization activity.
  • Tissue specificity
    Higher levels in osteoarthritic articular cartilage, aorta, uterus. Moderate expression in small intestine, heart, liver, bladder, ovary, stomach, and in the adrenal, thyroid, and mammary glands. Low expression in trachea, bone marrow, and lung. Co-localizes with TGFB1 in chondrocytes within osteoarthritic (OA) lesions of articular cartilage.
  • Involvement in disease
    Genetic variations in ASPN are associated with susceptibility to osteoarthritis type 3 (OS3) [MIM:607850]; also known as osteoarthritis of knee/hip. Osteoarthritis is a degenerative disease of the joints characterized by degradation of the hyaline articular cartilage and remodeling of the subchondral bone with sclerosis. Clinical symptoms include pain and joint stiffness often leading to significant disability and joint replacement. Note=Susceptibility to osteoarthritis is conferred by a triplet repeat expansion polymorphism. ASPN allele having 14 aspartic acid repeats in the N-terminal region of the protein (D14), is overrepresented relative to the common allele having 13 aspartic acid repeats (D13). The frequency of the D14 allele increases with disease severity. The D14 allele is also overrepresented in individuals with hip osteoarthritis.
    Defects in ASPN are a cause of susceptibility to intervertebral disk disease (IDD) [MIM:603932]. A common musculo-skeletal disorder caused by degeneration of intervertebral disks of the lumbar spine. It results in low-back pain and unilateral leg pain. Note=Susceptibility to intervertebral disk disease, particularly lumbar disk degeneration, is conferred by a triplet repeat expansion polymorphism. ASPN allele having 14 aspartic acid repeats in the N-terminal region of the protein (D14), is associated with the disorder in some populations (PubMed:18304494).
  • Sequence similarities
    Belongs to the small leucine-rich proteoglycan (SLRP) family. SLRP class I subfamily.
    Contains 11 LRR (leucine-rich) repeats.
    Contains 1 LRRNT domain.
  • Domain
    The LRR 5 repeat can inhibit BMP2-induced cytodifferentiation and may be involved in the interaction with BMP2 (By similarity). The repeats LRR 10, LRR 11 and LRR 12 are involved in binding type I collagen. The poly-Asp region is involved in binding calcium.
  • Post-translational
    modifications
    There is no serine/glycine dipeptide sequence expected for the attachment of O-linked glycosaminoglycans and this is probably not a proteoglycan. The O-linked polysaccharide on 54-Ser is probably the mucin type linked to GalNAc.
    The N-linked glycan at Asn-282 is composed of variable structures of GlcNAc, mannose, fucose, HexNAc and hexose.
  • Cellular localization
    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Database links
  • Alternative names
    • ASPN antibody
    • ASPN protein antibody
    • ASPN_HUMAN antibody
    • Asporin (LRR class 1) antibody
    • Asporin antibody
    • Asporin proteoglycan antibody
    • FLJ20129 antibody
    • LRR class 1 antibody
    • Periodontal ligament associated protein 1 antibody
    • Periodontal ligament-associated protein 1 antibody
    • PLAP 1 antibody
    • PLAP-1 antibody
    • PLAP1 antibody
    • SLRR 1C antibody
    • SLRR1C antibody
    • Small leucine rich protein 1C antibody
    see all

Images

  • All lanes : Anti-Asporin antibody (ab31303) at 1/1000 dilution

    All lanes : Human osteosarcoma cell line whole lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-goat IgG polyclonal at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 43 kDa
    Observed band size: 42 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocked with 5% milk for 1 hour at 27°C.

    Incubated with the primary antibody diluted in blocking buffer for 12 hours at 4°C.

    See Abreview

  • Anti-Asporin antibody (ab31303) at 0.3 µg/ml + Human Bone Marrow lysate (35µg protein in RIPA buffer)

    Predicted band size: 43 kDa
    Observed band size: 40 kDa why is the actual band size different from the predicted?



    Primary incubation was 1 hour. Detected by chemiluminescence.
  • ab31303 (4µg/ml) staining of paraffin embedded Human Colon. Steamed antigen retrieval with citrate buffer pH 6, HRP-staining. Shows textured staining in the cytoplasm of absorptive cells.
  • Ab31303 staining of ATDC5 cells (Panels A and C) and DAPI (panels B and D). Image shows staining of cytoplasm and on the cell surface. Data gratefuly received from Dr. Shiro Ikegawa, SNP Research Center, RIKEN, Japan.

References

This product has been referenced in:
  • Maccarana M  et al. Asporin-deficient mice have tougher skin and altered skin glycosaminoglycan content and structure. PLoS One 12:e0184028 (2017). Read more (PubMed: 28859141) »
  • Hurng JM  et al. Discontinuities in the human bone-PDL-cementum complex. Biomaterials 32:7106-17 (2011). ICC/IF ; Human . Read more (PubMed: 21774982) »
See all 4 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Osteosarcoma cell lines)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Osteosarcoma cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Jun 19 2015

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this peptide.

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 the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab134013 and ab31303.

The protocol looks good and also our lab could not find major difference to how they are testing. They are using more peptide (100ng/well)

There is only one concern: Standard ELISA plates will not be "sticky" enough for the small peptides and the peptide gets washed off and therefore there will be no signal.

We can recommend to use high bond plates (for example Costar).

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again.

Read More

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.

Read More

Answer


We will be adding the peptide to our catalogue, and I will inform you as soon as this has been done.

Read More

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 with the order number xxx with a new lot.

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.

Read More

Answer

Thank you for your enquiry.

I can confirm that this antibody was tested in IHC-P in the publication provided on the datasheet:

Ho SP et al. The biomechanical characteristics of the bone-periodontal ligament-cementum complex. Biomaterials 31:6635-46 (2010).
http://www.ncbi.nlm.nih.gov/pubmed/20541802?dopt=Abstract

The authors stained human periodontium tissue. This would be suitable as a positive control.

Alternatively, the Uniiprot protein database provides the following information on the expression of Asporin in the following tissues.Any high or moderate expressing tissuecould also be suitable as positive control:

'Higher levels in osteoarthritic articular cartilage, aorta, uterus. Moderate expression in small intestine, heart, liver, bladder, ovary, stomach, and in the adrenal, thyroid, and mammary glands. Low expression in trachea, bone marrow, and lung.'

I hope this information will be helpful to you. if you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for taking the time to contact us.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB, ELISA, IHC-P and ICC/IF with human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

Could you please clarify which application(s) you are using?

In order to provide protocol tips, and also obtain more information for our quality monitoring records, I have attached technical questionnaires. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you could also provide an image which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

Read More

Answer

Thank you for contacting Abcam. Below is a link to our general antigen-retrieval protocol, although the protocol may have to be optimized for each antibody: https://www.abcam.com/index.html?pageconfig=resource&rid=11488 However looking at some of the Abreviews, a customer has published a review saying that for ab6588 (Collagen VI) they did not need to perform antigen retrieval, I have included the reference for that paper: Stem Cells. 2008 Feb;26(2):570-9. Epub 2007 Dec 6. For ab31303 (Asporin), the following reference contains a protocol for staining: Biomaterials. 2010 Sep;31(25):6635-46. Epub 2010 Jun 11. The antibody ab74524 (COMP), has not been tested in IHC-P and so we cannot guarantee that it will work in this application, however I would suggest trying out a few different protocols with several antibody dilutions. Just because we do not state that it will work does not mean it will not work, we just cannot guarantee it. If there is anything I can do to be of further assistance then please do not hesitate to contact me. I hope the staining goes well.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (primary chondrocytes)
Loading amount
30 µg
Specification
primary chondrocytes
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Feb 28 2011

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Recombinant protein (from abnova Catalog # : H00054829-P01)
Specification
from abnova Catalog # : H00054829-P01
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Type
Direct

Abcam user community

Verified customer

Submitted Oct 06 2010

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

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