Anti-Aggrecan, MMP Cleaved, clone AF-28, Ascites Free (mouse monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
H, B, M, Porcine, R	WB, IH(P)	M	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

H, B, M, Porcine, R

WB, IH(P)

Purified

Monoclonal Antibody

Description
Catalogue Number	MAB19310-C
Description	Anti-Aggrecan Antibody, MMP Cleaved, clone AF-28, Ascites Free
Alternate Names	Aggrecan core protein, MMP-cleaved Cartilage-specific proteoglycan core protein, MMP-cleaved Chondroitin sulfate proteoglycan 1, MMP-cleaved Chondroitin sulfate proteoglycan core protein 1, MMP-cleaved CSPCP, MMP-cleaved
Background Information	Aggrecan core protein (UniProt P16112; also known as Cartilage-specific proteoglycan core protein, Chondroitin sulfate proteoglycan 1, Chondroitin sulfate proteoglycan core protein 1, CSPCP) is encoded by the ACAN (also known as AGC1, CSPG1, MSK16, SEDK) gene (Gene ID 176) in human. Aggrecan is the major extracellular matrix (ECM) proteoglycan in articular cartilage that, together with type II collagen, provides the cartilage with its mechanical properties of reversible compressibility. Aggrecan consists of two globular domains (G1 and G2) and an IGD (interglobular domain) sequence in between, followed by an extended region before the third globular domain (G3). The region between G2 and G3 is heavily substituted with negatively charged sGAG (sulfated glycosaminoglycan) and is further subdivided into the KS (keratan sulfate), CS1 (chondroitin sulfate region 1) and CS2 regions. Degradation of cartilage ECM, including aggrecan, is a hallmark in arthritic diseases and in joint injuries. Multiple aggrecan cleavage sites have been characterized, including those in G1 domain (by cathepsin K), IGD (by aggrecanases, calpain, cathepsins B and K), KS (by calpain and MMPs), CS1 (by calpain and MMPs), and CS2 (by aggrecanases, calpain, cathepsin D and MMPs). In addition, different MMPs exhibit differential affinities toward aggrecan, the IGD IPEN-FFGV site is cut by MMPs 1–3, 7–9, 12–16, 19 and 20, the CS1 region in bovine aggrecan is cut at multiple GVED-I/(L)SGL sites by MMP-3, the end of the CS2 region in bovine aggrecan is cut at RPAE-ARLE by MMPs 2, 3, 7 and 12. Aggrecan proteolysis in the IGD results in decreased tissue water-holding capacity due to a loss of sGAG-containing chains. On the other hand, aggrecan proteolysis in the CS1 and CS2 regions is a natural turnover process in mature cartilage and does not affect cartilage function.

Description

Catalogue Number

MAB19310-C

Description

Anti-Aggrecan Antibody, MMP Cleaved, clone AF-28, Ascites Free

Alternate Names

Aggrecan core protein, MMP-cleaved
Cartilage-specific proteoglycan core protein, MMP-cleaved
Chondroitin sulfate proteoglycan 1, MMP-cleaved
Chondroitin sulfate proteoglycan core protein 1, MMP-cleaved
CSPCP, MMP-cleaved

Background Information

Aggrecan core protein (UniProt P16112; also known as Cartilage-specific proteoglycan core protein, Chondroitin sulfate proteoglycan 1, Chondroitin sulfate proteoglycan core protein 1, CSPCP) is encoded by the ACAN (also known as AGC1, CSPG1, MSK16, SEDK) gene (Gene ID 176) in human. Aggrecan is the major extracellular matrix (ECM) proteoglycan in articular cartilage that, together with type II collagen, provides the cartilage with its mechanical properties of reversible compressibility. Aggrecan consists of two globular domains (G1 and G2) and an IGD (interglobular domain) sequence in between, followed by an extended region before the third globular domain (G3). The region between G2 and G3 is heavily substituted with negatively charged sGAG (sulfated glycosaminoglycan) and is further subdivided into the KS (keratan sulfate), CS1 (chondroitin sulfate region 1) and CS2 regions. Degradation of cartilage ECM, including aggrecan, is a hallmark in arthritic diseases and in joint injuries. Multiple aggrecan cleavage sites have been characterized, including those in G1 domain (by cathepsin K), IGD (by aggrecanases, calpain, cathepsins B and K), KS (by calpain and MMPs), CS1 (by calpain and MMPs), and CS2 (by aggrecanases, calpain, cathepsin D and MMPs). In addition, different MMPs exhibit differential affinities toward aggrecan, the IGD IPEN-FFGV site is cut by MMPs 1–3, 7–9, 12–16, 19 and 20, the CS1 region in bovine aggrecan is cut at multiple GVED-I/(L)SGL sites by MMP-3, the end of the CS2 region in bovine aggrecan is cut at RPAE-ARLE by MMPs 2, 3, 7 and 12. Aggrecan proteolysis in the IGD results in decreased tissue water-holding capacity due to a loss of sGAG-containing chains. On the other hand, aggrecan proteolysis in the CS1 and CS2 regions is a natural turnover process in mature cartilage and does not affect cartilage function.

Product Information
Format	Purified
Presentation	Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Product Information

Format

Purified

Presentation

Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Applications
Application	This Anti-Aggrecan Antibody, MMP Cleaved, clone AF-28, Ascites Free is validated for use in Western Blotting, Immunohistochemistry (Paraffin) for the detection of Aggrecan.
Key Applications	Western Blotting Immunohistochemistry (Paraffin)
Application Notes	Immunohistochemistry Analysis: A 1:50 dilution from a representative lot detected MMP-cleaved aggrecan in human cartilage tissue. Immunohistochemistry Analysis: A representative lot detected MMP-cleaved aggrecan in paraffin-embedded tibia sections from both wild-type mice and mice with decreased MMP-13 expression (Zhou, X., et al. (2010). Proc. Natl. Acad. Sci. U. S. A. 107(29):12919-12924). Western Blotting Analysis: A representative lot detected a higher level of MMP-generated FFGV fragments in osteoarthritis/osteoarthritic (OA) cartilage than in normal cartilage samples (Struglics, A., and Hansson, M. (2012). Biochem. J. 446(2):213-223). Western Blotting Analysis: A representative lot detected MMPs-digested G2 fragment from recombinant G1-G2, but not undigested, elastase- or trypsin-digested fragments (Mercuri, F.A., et al. (1999). J. Biol. Chem. 274(45):32387-32395). Western Blotting Analysis: A representative lot detected MMP-13-digested fragments from human, pig, bovine and rat, but not shark, aggrecan. Clone AF-28 did not detect undegraded aggrecan (Fosang, A.J., et al. (1996). FEBS Lett. 380(1-2):17-20). Western Blotting Analysis: A representative lot detected in vitro generated aggrecan fragments by MMPs, as well as various aggrecan fragments in arthritis patients-derived synovial fluids (Fosang, A.J., et al. (1995). Biochem. J. 310( Pt 1):337-343).

Applications

Application

This Anti-Aggrecan Antibody, MMP Cleaved, clone AF-28, Ascites Free is validated for use in Western Blotting, Immunohistochemistry (Paraffin) for the detection of Aggrecan.

Key Applications

Western Blotting
Immunohistochemistry (Paraffin)

Application Notes

Immunohistochemistry Analysis: A 1:50 dilution from a representative lot detected MMP-cleaved aggrecan in human cartilage tissue.
Immunohistochemistry Analysis: A representative lot detected MMP-cleaved aggrecan in paraffin-embedded tibia sections from both wild-type mice and mice with decreased MMP-13 expression (Zhou, X., et al. (2010). Proc. Natl. Acad. Sci. U. S. A. 107(29):12919-12924).
Western Blotting Analysis: A representative lot detected a higher level of MMP-generated FFGV fragments in osteoarthritis/osteoarthritic (OA) cartilage than in normal cartilage samples (Struglics, A., and Hansson, M. (2012). Biochem. J. 446(2):213-223).
Western Blotting Analysis: A representative lot detected MMPs-digested G2 fragment from recombinant G1-G2, but not undigested, elastase- or trypsin-digested fragments (Mercuri, F.A., et al. (1999). J. Biol. Chem. 274(45):32387-32395).
Western Blotting Analysis: A representative lot detected MMP-13-digested fragments from human, pig, bovine and rat, but not shark, aggrecan. Clone AF-28 did not detect undegraded aggrecan (Fosang, A.J., et al. (1996). FEBS Lett. 380(1-2):17-20).
Western Blotting Analysis: A representative lot detected in vitro generated aggrecan fragments by MMPs, as well as various aggrecan fragments in arthritis patients-derived synovial fluids (Fosang, A.J., et al. (1995). Biochem. J. 310( Pt 1):337-343).

Biological Information
Immunogen	KLH-conjugated linear peptide (FFGVGGEED-C) corresponding to a.a. 361-369 of human aggrecan core protein.
Epitope	N-terminus FFGVG neoepitope sequence.
Clone	AF-28
Concentration	Please refer to lot specific datasheet.
Host	Mouse
Specificity	Clone AF-28 is a cleavage-site-specific monoclonal antibody that recognizes polypeptides with N-terminal 345-FFGVG sequence (numbering based on mature form) found at the N-terminal end of MMP-digested aggrecan fragments. By immunoblotting, clone AF-28 specifically detected G2 fragments derived from an aggrecan G1-G2 substrate digested with stromelysin, collagenase, gelatinase, and matrilysin, but failed to detect the G2 fragments by elastase, trypsin, or cathepsin B cleavage. Competition studies confirmed that clone AF-28 does not react with peptides containing internal FFGVG sequence.
Isotype	IgG1κ
Species Reactivity	Human Bovine Mouse Porcine Rat
Species Reactivity Note	Human, Bovine, Mouse, Porcine, Rat.
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_037359
Gene Symbol	ACAN AGC1 CSPG1 MSK16 SEDK
Purification Method	Protein G purified.
UniProt Number	P16112
Molecular Weight	~200-260 kDa observed. Target fragment(s) appear larger than the calculated moleuclar weight(s) due to glycosylation. Uncharacterized band(s) may appear in some lysates.

Biological Information

Immunogen

KLH-conjugated linear peptide (FFGVGGEED-C) corresponding to a.a. 361-369 of human aggrecan core protein.

Epitope

N-terminus FFGVG neoepitope sequence.

Clone

AF-28

Concentration

Please refer to lot specific datasheet.

Host

Mouse

Specificity

Clone AF-28 is a cleavage-site-specific monoclonal antibody that recognizes polypeptides with N-terminal 345-FFGVG sequence (numbering based on mature form) found at the N-terminal end of MMP-digested aggrecan fragments. By immunoblotting, clone AF-28 specifically detected G2 fragments derived from an aggrecan G1-G2 substrate digested with stromelysin, collagenase, gelatinase, and matrilysin, but failed to detect the G2 fragments by elastase, trypsin, or cathepsin B cleavage. Competition studies confirmed that clone AF-28 does not react with peptides containing internal FFGVG sequence.

Isotype

IgG1κ

Species Reactivity

Human
Bovine
Mouse
Porcine
Rat

Species Reactivity Note

Human, Bovine, Mouse, Porcine, Rat.

Antibody Type

Monoclonal Antibody

Entrez Gene Number

NP_037359

Gene Symbol

ACAN
AGC1
CSPG1
MSK16
SEDK

Purification Method

Protein G purified.

UniProt Number

P16112

Molecular Weight

~200-260 kDa observed. Target fragment(s) appear larger than the calculated moleuclar weight(s) due to glycosylation. Uncharacterized band(s) may appear in some lysates.

Product Usage Statements
Quality Assurance	Evaluated by Western Blotting in human chondrocyte tissue lysate. Western Blotting Analysis: 0.5 µg/mL of this antibody detected MMP-cleaved aggrecan in 10 µg of human chondrocyte tissue lysate.
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Quality Assurance

Evaluated by Western Blotting in human chondrocyte tissue lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected MMP-cleaved aggrecan in 10 µg of human chondrocyte tissue lysate.

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Stable for 1 year at 2-8°C from date of receipt.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at 2-8°C from date of receipt.

Packaging Information
Material Size	100 μg

Packaging Information

Material Size

100 μg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Anti-Aggrecan, MMP Cleaved, clone AF-28, Ascites Free (mouse monoclonal)

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