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Aggrecan ELISA Kit (GWB-DE7FB7)



1x96 well plate
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Product Name: PG-EASIA

Category: Aggrecan

Sample Type: Synovial Fluids And Serum

Sensitivity: Bone Metabolism

Description: An immunoenzymetric assay for the quantitative measurement of human Aggrecan (PG) in synovial fluid, serum and cell culture

Intended Use: Aggrecan (PG) is the predominant proteoglycan species in articular cartilage. It is composed of a core protein of 210 kDa to which over 100 chondroitin sulfate chains, about 20-50 keratan sulfate chains and O-linked as well as N-linked oligosaccharides are covently attached. The core protein contains three distinct globular domains (G1-G3). G1 is at the amino terminus, separated by a short extended segment from G2, while G3 is at the carboxy terminal end. The G1 amino terminal region can interact noncovalently with hyaluronic acid (HA) and has then be termed the hyaluronic acid binding region (HABR). A link protein interact with both the G1 region and the HA to stabilize this interaction. PG is produced by chondrocytes, and its production is regulated by cytokines and growth factors such as IL1?, TNF?, IGF1 or TGF?. In extracellular matrix, as many as 200 aggrecan molecules can bind to one single HA molecule to form an aggregate (MW : 5 10^7 to 5 10^8 ).
An imbalance in the synthesis and degradation of the matrix components is a common feature of both osteoarthritis and rheumatoid arthritis. The loss of PG and other matrix components from the cartilage leads to destruction of the tissue, causing complete deterioration of the articular surface. Several recent publications suggest that the PG and PG fragments released in synovial fluid and serum during the degradation process might serve as markers of the metabolic changes in diseased cartilage. Cell culture is a commonly used procedure for the study of cartilage metabolism. The measurement of PG and other matrix components in culture supernatants and cellular contents can assist analysis of the effects of cytokines, growth factors, drugs and potential chondroprotective substances on the cartilage homeostasy. GenWay Biotech has developped a ELISA for the measurement of human aggrecan to aid the study of this important cartilage constituent. The PG-EASIA is convenient, highly specific, and allow accurate measurement of PG in synovial fluid, serum and culture supernatant This assay is a new interesting tool for the exploration of the cartilage metabolism.

Principles of the assay: The PG-EASIA is a solid phase Enzyme Amplified Sensitivity Immunoassay (EASIA) performed on microtiter plate. The assay is based on an oligoclonal system in which a blend of monoclonal antibodies (MAbs) directed against distinct epitopes of PG are used. Antibody-producing cells are immortilized using the myeloma cell fusion method of Kohler and Milstein. A hybridoma cell is produced which secretes specific homogeneous antibodies. The use of a number of distinct MAbs avoids hyperspecificity and allows highly sensitive assays with extended standard range and short incubation time. Standards or samples containing PG react with capture monoclonal antibodies (MAbs 1) coated on the microtiter well and with a monoclonal antibody (MAb 2) labelled with horseradish peroxidase (HRP). After an incubation period allowing the formation of a sandwich : coated MAbs 1 - PG - MAb 2 - HRP, the microtiter plate is washed to remove unbound enzyme labelled antibodies. Bound enzyme-labelled antibodies are measured through a chromogenic reaction. Chromogenic Solution (TMB+H2O2) is added and incubated. The reaction is stopped with the addition of Stop Solution (HCl) and the microtiter plate is then read at the appropriate wavelength. The amount of substrate turnover is determined colourimetrically by measuring the absorbance which is proportional to the PG concentration. A standard curve is plotted and PG concentrations in a sample is determined by interpolation from the standard curve. The use of the EASIA Reader (linearity up to 3 OD units) and a sophisticated data reduction method (polychromatic data reduction) result in high sensitivity in the low range and in an extended standard range.

Storage and Stability: A. UNOPENED vials
Store the unopened vials at 2C to 8C. All kit components are stable until the expiry date printed on the labels.
B. OPENED vials
1. The Conjugate vial must be stored at 2 to 8C.
2. The reconstituted Standards and Controls are stable for 1 day at 2C to 8C. Aliquots held for longer periods of time should be frozen, a maximum of two times, at -20C (maximum 2 months) or at -70C for longer storage (until expiration date).
3. Store the unused strips at 2C to 8C in the sealed bag containing the dessiccant until expiration date.
4. The Wash Solution Concentrate is stable at room temperature until expiration date. In order to avoid washerhead obstructions, it is recommended to prepare a fresh diluted Wash Solution each day.

Product Name: PG-EASIA

Function: This proteoglycan is a major component of extracellular matrix of cartilagenous tissues. A major function of this protein is to resist compression in cartilage. It binds avidly to hyaluronic acid via an N-terminal globular region.

Subunit: Interacts with FBLN1 (By similarity).

Subcellular Location: Secreted, extracellular space, extracellular matrix (By similarity).

Tissue Specificity: Restricted to cartilages.

Developmental Stage: Expression was detected in chondrocytes throughout the developing skeleton.

Domain: Two globular domains, G1 and G2, comprise the N-terminus of the proteoglycan, while another globular region, G3, makes up the C-terminus. G1 contains Link domains and thus consists of three disulfide-bonded loop structures designated as the A, B, B\' motifs. G2 is similar to G1. The keratan sulfate (KS) and the chondroitin sulfate (CS) attachment domains lie between G2 and G3.

Ptm: Contains mostly chondroitin sulfate, but also keratan sulfate chains, N-linked and O-linked oligosaccharides. The release of aggrecan fragments from articular cartilage into the synovial fluid at all stages of human osteoarthritis is the result of cleavage by aggrecanase.

Disease: Defects in ACAN are the cause of spondyloepiphyseal dysplasia type Kimberley (SEDK) [MIM:608361]. Spondyloepiphyseal dysplasias are a heterogeneous group of congenital chondrodysplasias that specifically affect epiphyses and vertebrae. The autosomal dominant SEDK is associated with premature degenerative arthropathy.

Similarity: Belongs to the aggrecan/versican proteoglycan family.

Similarity: Contains 1 C-type lectin domain.

Similarity: Contains 1 EGF-like domain.

Similarity: Contains 1 Ig-like V-type (immunoglobulin-like) domain.

Similarity: Contains 4 Link domains.

Similarity: Contains 1 Sushi (CCP/SCR) domain.

Additional Information

Name Aggrecan ELISA Kit (GWB-DE7FB7)
Related Product Names Aggrecan ELISA Kit; Cartilage-specific proteoglycan core protein; CSPCP; Chondroitin sulfate proteoglycan core protein 1 AGC1; CSPG1; MSK16AggrecanPG
Swissprot ID P16112
NCBI Acc Number NP_001126.2
Molecular Weight 250193
Datasheets/Manuals Download manual for GWB-DE7FB7
Intended Use Research Use Only