Immunofluorescence analysis of Aggrecan G3 was performed using 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Aggrecan G3 Rabbit Polyclonal Antibody (Product # PA1-1745) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization in the extracellular matrix. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Dog, Human, Mouse, Rat|
|Published species reactivity||Bacteria, Fish, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C D(2277) G H P M Q F E N W R P N Q P D N(2293) of human aggrecan.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 6 publications below|
PA1-1745 detects human, rat, mouse, bovine, and canine aggrecan G3.
PA1-1745 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~350 kDa protein representing the aggrecan G3 domain on the full length core.
The PA1-1745 immunogen is a synthetic peptide corresponding to residues C D(2277) G H P M Q F E N W R P N Q P D N(2293) of human aggrecan.. This antibody is also referred to as anti-G3.
Aggrecan is a member of large, aggregating proteoglycans (also including, versican, brevican and neurocan) found in articular cartilage. Aggrecan is composed of three major domains: G1, G2, and G3. Between the G1 and G2 domains there is an interglobulin region (IGD). The IGD region is the major site of cleavage by specific proteases like metalloproteinases (MMP and quote;s) and aggrecanase. Aggrecan cleavage has been associated with a number of degenerative diseases including rheumatoid arthritis and osteoarthritis.
The G3 domain is a complicated region produced by alternative splicing of exons in post-transcriptional processes. The human G3 domain consists of three modules and articular cartilage contains a mixture of the alternatively spliced G3 modules. G3 has been shown to facilitate glycosaminoglycan (GAG) attachment and overall aggrecan processing. The G3 domain is central to aggrecan turnover and function in cartilage.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Biochemical and atomic force microscopic characterization of salmon nasal cartilage proteoglycan.
PA1-1745 was used in western blot to study salmon nasal cartilage proteoglycan using a combination of biochemistry and atomic force microscopy
|Kakizaki I,Mineta T,Sasaki M,Tatara Y,Makino E,Kato Y||Carbohydrate polymers (103:538)||2014|
Aggrecanase cleavage in juvenile idiopathic arthritis patients is minimally detected in the aggrecan interglobular domain but robust at the aggrecan C-terminus.
PA1-1745 was used in western blot to study the pattern of aggrecan degradation in patients with idiopathic juvenile arthritis
|Struglics A,Lohmander LS,Last K,Akikusa J,Allen R,Fosang AJ||Arthritis and rheumatism (64:4151)||2012|
MMP proteolysis of the human extracellular matrix protein aggrecan is mainly a process of normal turnover.
PA1-1745 was used in western blot to study the MMP-mediated proteolysis of extracellular matrix aggrecan
|Struglics A,Hansson M||The Biochemical journal (446:213)||2012|
Identification of proteoglycan from salmon nasal cartilage.
PA1-1745 was used in western blot to investigate the structural composition of proteoglycan in salmon nasal cartilage
|Kakizaki I,Tatara Y,Majima M,Kato Y,Endo M||Archives of biochemistry and biophysics (506:58)||2011|
Calpain is involved in C-terminal truncation of human aggrecan.
PA1-1745 was used in western blot to study the role oh calpain in the truncation of human aggrecan.
|Struglics A,Hansson M||The Biochemical journal (430:531)||2010|
A comparison of different purification methods of aggrecan fragments from human articular cartilage and synovial fluid.
PA1-1745 was used in western blot to evaluate the effectiveness of various purification methods of aggrecan purification from human cartilage and synovial fluid
|Struglics A,Larsson S||Matrix biology : journal of the International Society for Matrix Biology (29:74)||2010|