Aggrecan Antibody (BC-3)

• Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Eq, Fe, Gp, Rb, Sh
• Applications: WB, ELISA, ICC/IF, IHC, IHC-Fr, IHC-P
• Clone: BC-3
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Aggrecan Antibody (BC-3) Summary

• Immunogen: ARGSV... synthetic peptide conjugate.
• Localization: Secreted
• Marker: Chondrocyte Marker
• Specificity: Recognizes the aggrecanase (ADAMTS-1, -4 & -5)-generated N-terminal neoepitope ARG after cleavage between amino acids EGE and ARG within the interglobular domain of aggrecanase-catabolised aggrecan.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: ACAN
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:100
  • ELISA 1:100-1:2000
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
• Application Notes: This Aggrecan antibody is useful in Immunohistochemistry, ELISA and Western blot. With IHC the antibody works in formalin- or paraformaldehyde-fixed paraffin embedded sections as well as either alcohol-fixed frozen sections or un-fixed snap-frozen sections. In Western blot the antibody detects a variety of epitopes between 50 and 250 kDa. Samples must be deglycosylated using 0.01 Units Chondroitinase ABC (Sigma), 0.01 Units Keratanase (Seikagaku) and 0.0001 Units Keratanase II (Seikagaku) per 10ug S-GAG of non-deglycosylated aggrecan for optimal epitope recognition. See Little et al. and Caterson et al. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 27185069).
• Theoretical MW: 250 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Cross-reacts with Human, Cow, Cat, Dog, Guinea pig, Horse, Pig, Rabbit, Rat and Sheep. Mouse reactivity reported in scientific literature (PMID: 27185069). Not yet tested in other species.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein A or G purified

Alternate Names for Aggrecan Antibody (BC-3)

• ACAN
• AGC1
• AGC1SEDK
• aggrecan core protein
• Aggrecan
• Cartilage-specific proteoglycan core protein
• Chondroitin sulfate proteoglycan 1
• Chondroitin sulfate proteoglycan core protein 1
• CSPCP
• CSPG1
• CSPG1aggrecan 1
• CSPGCP
• large aggregating proteoglycan
• MSK16
• MSK16AGCAN
• SEDK

Background

The Aggrecan degradation product usually remains within the tissue still complexed to hyaluronan and link protein. Its release from the cartilage usually signals that there has been extensive catabolism of aggrecan, which allows large complexes containing this metabolite to be released from the tissue. Aggrecan degradation products can be found in the synovial fluids of patients with degenerative joint disease.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Aggrecan Antibody (NB600-504)(12)

Publications using NB600-504

• Si HB, Yang TM, Li L et al. miR-140 Attenuates the Progression of Early-Stage Osteoarthritis by Retarding Chondrocyte Senescence Mol Ther Nucleic Acids Nov 9 2019 [PMID: 31790972] (WB, IHC-P, Human, Rat)
• Cheng B, Liu Y, Zhao Y et al. The role of anthrax toxin protein receptor 1 as a new mechanosensor molecule and its mechanotransduction in BMSCs under hydrostatic pressure Sci Rep Sep 2 2019 [PMID: 31477767] (WB, Rat)
• Liu Z, Zhang Z, Zhang A et al. Osteogenic protein-1 alleviates high glucose microenvironment-caused degenerative changes in nucleus pulposus cells Biosci. Rep. Apr 30 2019 [PMID: 30872412] (ICC/IF, Rat)
• Chen L, Liu G, Li W, Wu X. Chondrogenic differentiation of bone marrow-derived mesenchymal stem cells following transfection with Indian hedgehog and sonic hedgehog using a rotary cell culture system Cell Mol Biol Lett Dec 1 2019 [PMID: 30858866] (WB, Rabbit)
• Jia Jun, Wang Shan-Zheng, Ma Liang-Yu et al. The Differential Effects of Leukocyte-Containing and Pure Platelet-Rich Plasma on Nucleus Pulposus-Derived Mesenchymal Stem Cells: Implications for the Clinical Treatment of Intervertebral Disc Degeneration. Stem Cells International 2018 Oct 23 [PMID: 30425747] (ICC/IF, WB, Rabbit)
• Yang D, Zhu D, Zhu S et al. 17 beta-Estradiol/extrogen receptor beta alleviates apoptosis and enhances matrix biosynthesis of nucleus pulposus cells through regulating oxidative damage under a high glucose condition Biomedicine & Pharmacotherapy Nov 1 2018 [PMID: 30257311] (ICC/IF, Rat)
• Han ME, Kim SH1, Kim HD et al. Extracellular Matrix-based Cryogels for Cartilage Tissue Engineering. Int J Biol Macromol. [PMID: 27185069] (IHC, Mouse)
• Bouhenni R, Hart M, Al-Jastaneiah S et al. Immunohistochemical expression and distribution of proteoglycans and collagens in sclerocornea. Int Ophthalmol. 2013 Jan [PMID: 23325424] (IHC-P, Human)
• Yurube T, Takada T, Suzuki T, Kakutani K, Maeno K, Doita M, Kurosaka M, Nishida K. Rat tail static compression model mimics extracellular matrix metabolic imbalances of matrix metalloproteinases, aggrecanases, and tissue inhibitors of metalloproteinases in intervertebral disc degeneration. Arthritis Res Ther;14(2):R51. 2012 Mar 6. [PMID: 22394620] (IHC, Rat)
• Swearingen CA, Chambers MG, Lin C et al. A short-term pharmacodynamic model for monitoring aggrecanase activity: injection of monosodium iodoacetate (MIA) in rats and assessment of aggrecan neoepitope release in synovial fluid using novel ELISAs. Osteoarthritis Cartilage 2010 Sep [PMID: 20633676] (WB, Bovine, Human, Rat)
• Little CB, Hughes CE, Curtis CL, Janusz MJ, Bohne R, Wang-WeigS, Taiwo YO, Mitchell PG, Otterness IG, Flannery CR, Caterson B. Matrix metalloproteinases are involved in C-terminal and interglobular domain processing of cartilage aggrecan in late stage cartilage degradation. Matrix Biol;21(3):271-88. 2002 Apr. [PMID: 12009333]
• Ma S, Xie Z, Yan G et al. Effect of Acupotomy on FAK-PI3K signaling pathways in KOA rabbit articular cartilages Evidence-Based Complementary and Alternative Medicine 2017 Aug 28 (Rabbit)

FAQs for Aggrecan Antibody (NB600-504). (Showing 1 - 3 of 3 FAQs).

1. Our customer would like to confirm the predicted MW by using NB600-504 to detect Aggrecan. According to info from http://www.uniprot.org/uniprot/P16112, the MW of the target protein may be between 239 to 250 kDa. But this antibody can also be used with ELISA and Western blot, where it detects a variety of epitopes between 50 and 250 kDa is described. May we have the explanation?
   • There are two isoforms of aggrecan, and this antibody can detect either one or both of them. It depends on which isoforms a particular cell type/tissue/species expresses.
2. Do you know if this antibody has been used with immunofluorescence?
   • We have not yet validated this antibody for use in immunofluorescence. In instances such as this, we recommend our Innovators Reward Program. Under the terms and conditions of this program, we would be happy to provide you with a credit of equal or lesser value, in exchange for your online review of this product including an image of either positive or negative staining for testing in a new species and/or application.
3. On behalf of a customer I have a question about antibody NB600-504 anti-ACAN. The note on the datasheet says Samples must be deglycosylated using 0.01 Units Chondroitinase ABC (Sigma), 0.01 Units Keratanase (Seikagaku) and 0.0001 Units Keratanase II (Seikagaku) per 10ug S-GAG of non-deglycosylated aggrecan for optimal epitope recognition. Do you really mean to say deglycosylated? Are not those reagents actually digesting the aggrecan? Do they also deglycosylate the protein?
   • Aggrecan is a highly glycosylated protein and this can interfere with the antibody binding to and recognizing the epitope. These enzymes will deglycosylate the protein and expose the epitope for recognition by the antibody

Bioinformatics

• Gene Symbol: ACAN
• Entrez:
  • 176(Human)
  • 58968(Rat)
• Uniprot:
  • P16112()