Aggrecan was detected in immersion fixed rat mesenchymal stem cells differentiated to chondrocytes using Goat Anti-Human Aggrecan G1-IGD-G2 Domains Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1220) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conju-gated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Aggrecan in Human Cartilage.

Aggrecan was detected in immersion fixed paraffin-embedded sections of human cartilage using 15 µg/mL Goat Anti-Human Aggrecan G1-IGD-G2 Domains Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1220) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Porcine Aggrecan by Immunocytochemistry/ Immunofluorescence

In vitro differentiation capacity of young and old BM-MSCs: (A) Representative micrographs from each experimental group showing morphological change after 21 days of differentiation. (B) Results of adipogenic, osteogenic and chondrogenic differentiation. Representative micrographs showing anti-FABP4, Oil Red O, anti-osteocalcin, Alizarin Red-S and anti-aggrecan staining in cultured BM-MSCs from each experimental group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/21992089), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Aggrecan G1-IGD-G2 Domains Antibody

Application

Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed rat mesenchymal stem cells differentiated into chondrocytes

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human cartilage

Western Blot

0.1 µg/mL
Sample: Recombinant Human Aggrecan G1-IGD-G2 Domains (Catalog # 1220-PG)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Aggrecan

Aggrecan, also known as aggrecan 1, chondroitin sulfate proteoglycan, and large aggregating proteoglycan, is encoded by the AGC1 gene with gene aliases of SEDK; CSPG1; MSK16; CSPGCP. As the key component of the cartilage extracellular matrix, aggrecan hydrates the collagen network and provides cartilage with its properties of compressibility and elasticity. Maintenance of aggrecan content is therefore critical to the function of the tissue and aggrecan degradation is an important factor in the erosion of articular cartilage in arthritic diseases (2). The deduced amino acid sequence of human aggrecan core protein consists of 2415 resides and predicts a signal peptide and domains of G1, IGD, G2, KS, CS-1, CS-2, and G3 (3). Two globular domains, G1 and G2, comprise the N-terminus of the proteoglycan and also contain link domains. The third globular domain, G3, corresponds to the C-terminus. The keratan sulfate (KS) and the chondroitin sulfate (CS) attachment domains are between G2 and G3. With KS and CS attached to the 250 kDa core protein, aggrecan monomers exist as a 1,000-2,000 kDa molecule. In addition, aggrecan monomers interact with hyaluronan through their G1 domain, resulting in larger aggregates containing 10-100 aggrecan monomers on a hyaluronan backbone (2).

Aggrecan can be cleaved by MMPs and ADAMTSs at the Asn360-Phe361 and Glu392-Ala393 bond in the IGD (residues are numbered based on Accession # NP_037359), respectively (2). Inhibition of ADATMS4 and ADAMTS5 cleavage prevents aggrecan degradation in osteoarthritic cartilage, while mice with aggrecan resistant to MMP cleavage do not accumulate aggrean and develop normally (2, 4). Consisting of the G1, IGD and G2 domains, rhAggrecan can be used as a protein substrate for MMPs and ADAMTSs and in binding assays involving hyaluronan.

References

Doege. K.J. et al. (1991) J. Biol. Chem. 266:894.
Malfait, A-M. et al. (2002) J. Biol. Chem. 277:22201.
Caterson, B. et al. (2000) Matrix Biol. 19:333.
Little, C.B. et al. (2005) Mol. Cel. Biol. 25:3388.

Alternate Names

ACAN, AGC1, CSPG1, CSPGCP, MSK16, SEDK

Entrez Gene IDs

176 (Human)

Gene Symbol

ACAN

UniProt

NP_037359

Additional Aggrecan Products

Product Documents for Human Aggrecan G1-IGD-G2 Domains Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human Aggrecan G1-IGD-G2 Domains Antibody

For research use only

Citations
Reviews

Human Aggrecan G1-IGD-G2 Domains Antibody

R&D Systems, part of Bio-Techne | Catalog # AF1220

Key Product Details

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

View all Aggrecan Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human Aggrecan G1-IGD-G2 Domains Antibody

Aggrecan in Rat Mesenchymal Stem Cells.