Immunofluorescence analysis of ADAMTS9 was performed using 70% confluent log phase SH-SY5Y 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 ADAMTS9 Rabbit Polyclonal Antibody (Product # PA1-1760) 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 extracellular matrix. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C F D G K H F N I N G L L P N of human ADAMTS9.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||0.5 ug/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.
PA1-1760 detects ADAMTS9 from human samples.
PA1-1760 has been successfully used in Western blotting procedures.
PA1-1760 immunogen is a synthetic peptide corresponding to residues C F D G K H F N I N G L L P N of human ADAMTS9. This polyclonal antibody has been referred to as JSCFDG.
ADAMTS proteases are secreted enzymes containing a prometalloprotease domain of the reprolysin type. The ADAMTS proteases function in processing of procollagens and von Willebrand factor as well as catabolism of aggrecan, versican and brevican. They have been demonstrated to have important roles in connective tissue organization, coagulation, inflammation, arthritis, angiogenesis and cell migration. The immunizing peptide sequence is 100% conserved between human, mouse and rat. ADAMTS9 is a member of the ADAMs family of proteinases with Thrombospondin motifs. ADAMTS9 is closest in homology to ADAMTS20, sharing 54% overall identity, and like ADAMTS20 ADAMTS9 gas a GON-like domain at the carboxyterminal end. The GON domain at the carboxyterminal end is similar to the GON-1 protein in C. elegans, mutations of which lead to defective gonadal development. ADAMTS9 is expressed in ovary and testis, but little is known about the role of ADAMTS9 in reproductive organs. ADAMTS9 is also expressed in the heart, placenta, lung, skeletal tissue, and pancreas, so the protein must have wider functions than just gonadal development. ADAMTS9, like ADAMTS20, has a total of 15 thrombospondin-like domains.
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