Immunofluorescence analysis of ADAMTS4 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 ADAMTS4 Rabbit Polyclonal Antibody (PA1-1749A) 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 conjugate (A27034) 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 (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 cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Human, Mouse, Pig|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C V(394) M A H V D P E E P(403) of mouse ADAMTS4.|
|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)||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.
PA1-1749A detects ADAMTS4 from porcine, bovine, mouse and human samples. PA1-1749A is specific for ADAMTS4 and shows no cross-reactivity with other ADAMTS family members.
PA1-1749A has been successfully used in Western blot procedures. By Western blot, this antibody detects ADAMTS4 at ~68, ~53 and 30 kDa protein representing ADAMTS4 and its proteolytic fragments. Depending on the extent of C-terminal processing, several smaller bands may also be seen using this antibody.
PA1-1749A immunogen is a synthetic peptide corresponding to residues C V(394) M A H V D P E E P(403) of mouse ADAMTS4. PA1-1749A immunizing peptide (Cat. # PEP-234) is available for use in neutralization and control procedures. This polyclonal antibody has been referred to as: JSCVMA. PA1-1749A has been used in combination with PA1-1750 for detection of all ADAMTS4 processed forms.
A disintegrin-like and metalloproteinase thrombospondin type 1 motif 4 (ADAMTS4 or aggrecanase 1) is a member of the ADAMTS family of multidomain proteins that has demonstrated metalloproteinase functions. The ADAMTS4 protein contains a prohormone convertase cleavage site, thrombospondin-1 like domain, cysteine-rich domain, and a propeptide domain. ADAMTS4 activity had been shown to be blocked by TIMP-1, 2, 3, and 4.
ADAMTS4 plays an important role in the cleavage of aggrecan, versican and brevican. Studies have determined that aggrecan is cleaved within the interglobular domain of aggrecan at sites Glu(373)-Ala(374) and Phe(342)-Glu(373). Aggrecan degradation has been linked to the development of rheumatoid arthritis, osteoarthritis, retinal diseases, Alzheimer and quote;s and other neurodegenerative diseases.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
ADAMTS-4 and biglycan are expressed at high levels and co-localize to podosomes during endothelial cell tubulogenesis in vitro.
PA1-1749A was used in western blot to study the expression of ADAMTS-4 and biglycan during endothelial cell tubulogenesis using an in vitro model
|Obika M,Vernon RB,Gooden MD,Braun KR,Chan CK,Wight TN||The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (62:34)||2014|
The expression of a disintegrin and metalloproteinase with thrombospondin motifs 4 in human macrophages is inhibited by the anti-atherogenic cytokine transforming growth factor-ß and requires Smads, p38 mitogen-activated protein kinase and c-Jun.
PA1-1749A was used in western blot to investigate the involvement of ADAMTS-4 in the anti-atherogenic effect of transforming growth factor
|Salter RC,Arnaoutakis K,Michael DR,Singh NN,Ashlin TG,Buckley ML,Kwan AP,Ramji DP||The international journal of biochemistry and cell biology (43:805)||2011|
The anti-atherogenic cytokine interleukin-33 inhibits the expression of a disintegrin and metalloproteinase with thrombospondin motifs-1, -4 and -5 in human macrophages: Requirement of extracellular signal-regulated kinase, c-Jun N-terminal kinase and phosphoinositide 3-kinase signaling pathways.
PA1-1749A was used in immunohistochemistry - paraffin section to investigate the effect of IL-33 on the expression of ADAMTS-1, -4 and -5 in human macrophages and atherosclerotic lesions
|Ashlin TG,Buckley ML,Salter RC,Johnson JL,Kwan AP,Ramji DP||The international journal of biochemistry and cell biology (46:113)||2014|