Invitrogen

ADAMTS4 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

14 Published Figures

10 References

View all (23) ADAMTS4 antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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ADAMTS4 Antibody in Immunocytochemistry (ICC/IF) — 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 (Product # 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 (Product # 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 (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 cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

ADAMTS4 Antibody in Western Blot (WB) — Fig1: Western blot analysis was performed using membrane enriched extracts (30 µg lysate) of SH-SY5Y (Lane 1), U-87MG (Lane 2) and Hep G2 (Lane3). Fig2: Western blot analysis was performed using 10 µL conditioned media of SH-SY5Y (Lane 1), U-87 MG (Lane 2) and Hep G2 (Lane3).The blots were probed with ADAMTS4 Rabbit polyclonal Antibody (Product # PA1-1749A, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). In Fig1. 68, 53 and 30 kDa bands corresponding to ADAMTS4 were observed across the cell lines tested. Along with the desired bands, non-specific bands were observed across cell lines. In Fig.2, ~ 68 kDa band corresponding to ADAMTS4 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

ADAMTS4 Antibody in Immunohistochemistry (IHC) — Confirmation of the specific binding of the ADAMTS4-specific probe. a From left to right: In vivo MRI before and after administration of the ADAMTS4-specific probe in one male mouse 4 weeks following aneurysms induction was highly comparable to corresponding ex vivo MRI scan of the abdominal aorta and ex vivo immunofluorescence staining (ADAMTS4 stain) and Elastica van Gieson (EvG) staining. b Intravenous administration of the fluorescent variant of the ADAMTS4-specific probe resulted in a strong fluorescence of the aneurysmal wall that was clearly co-localized with areas of ADAMTS4 expression using immunofluorescence staining (white arrows). c The competition experiment ( n = 3 mice) showed a significant decrease in the contrast-to-noise-ratio compared to the administration of the gadolinium-labeled ADAMTS4-specific probe alone, confirming the specific binding of the probe ( P = 0.0025). Data are presented as mean values +- SEM. d , e Using the negative control probe Cys*-Phe-His-Pro-Tyr-Cys*-linker (DOTA-Gd) with a similar size to the ADAMTS4-specific probe, no significant increase in contrast-to-noise ratio (CNR) was measured in vivo in the aortic wall of ApoE -/- mice ( n = 6). The application of the ADAMTS4-specific probe resulted in a strong and significant increase in CNR in the vessel wall as well in the thrombus. * P = 0.08; ** P = 0.04; Data are presented as mean values +- SEM. FLP Fluorescein-labeled probe, Scale bars indicate 100 um. * indicate the aortic lumen; #

ADAMTS4 Antibody — ADAMTS-4 cleaves Reelin at both the N- and C-terminus. (A-D, F) Anti-Reelin (G10, N-terminal antibody) immunoblots (IB). (A,B) Samples from Reelin-expressing HEK293 cells were collected after 12 hours incubation. (A) Metal chelator (PO, 50 µM), catechins (EGCG, 150 µM; ECG, 150 µM), and picetanol (PIC, 60 µM) inhibit Reelin cleavage. (B) Expression of TIMP-3, -1, and α2M, but not TIMP-4, inhibit Reelin cleavage. (C) Recombinant ADAMTS-4 (10 ng/µl) processes Reelin at both cleavage sites. Short vertical line at the bottom of the blot denotes that the last lanes, from the same blot, were joined for visual presentation. (D) Recombinant TIMP-3 (1.5, 3, 6, 12 ng/µl) and α2M (44, 72, 88,100 ng/µl) inhibit the proteolytic action of ADAMTS-4 on full-length Reelin. (E) Anti-ADAMTS-4 immunoblots (IB) showing reduction of ADAMTS-4 protein levels in HeLa cells transfected with two different ADAMTS-4 shRNA constructs, but not with scrambled shRNA nor with empty vector. (F) ADAMTS-4 shRNA transiently inhibited Reelin cleavage in Reelin-expressing HeLa cells. The frame emcompasses overexposed lanes from the blot on the right (asterisks) for better visualization of the Reelin fragments.

Product Details

PA1-1749A

Applications

Tested Dilution

Publications

Western Blot (WB)

1-3 µg/mL

View 5 publications 5 publications

Immunohistochemistry (IHC)

View 3 publications 3 publications

Immunohistochemistry (Paraffin) (IHC (P))

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

2 µg/mL

View 1 publication 1 publication

Product Specifications

Species Reactivity

Bovine, Human, Mouse, Pig

Published species

Human

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide corresponding to residues C V(394) M A H V D P E E P(403) of mouse ADAMTS4.

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1.5 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS with 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_1070004

Product Specific Information

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.

Target Information

This gene encodes a member of the ADAMTS protein family. Members of the family share several distinct protein modules, including a propeptide region, a metalloproteinase domain, a disintegrin-like domain, and a thrombospondin type 1 motif. Individual members of this family differ in the number of C-terminal TS motifs, and some have unique C-terminal domains. The enzyme encoded by this gene lacks a C-terminal TS motif. It is responsible for the degradation of aggrecan, a major proteoglycan of cartilage, and brevican, a brain-specific extracellular matrix protein. The cleavage of aggrecan and brevican suggests key roles of this enzyme in arthritic disease and in the central nervous system, potentially, in the progression of glioma.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Identification of Adamts4 as a novel adult cardiac injury biomarker with therapeutic implications in patients with cardiac injuries.

Scientific reports

Khanam R,Sengupta A,Mukhopadhyay D,Chakraborty S

Published figure using ADAMTS4 polyclonal antibody (Product # PA1-1749A) in Western Blot

Tue Jun 14 00:00:00 EDT 2022

Identification of Adamts4 as a novel adult cardiac injury biomarker with therapeutic implications in patients with cardiac injuries.

Scientific reports

Khanam R,Sengupta A,Mukhopadhyay D,Chakraborty S

Published figure using ADAMTS4 polyclonal antibody (Product # PA1-1749A) in Western Blot

Tue Jun 14 00:00:00 EDT 2022

ADAMTS4-specific MR probe to assess aortic aneurysms in vivo using synthetic peptide libraries.

Nature communications

Kaufmann JO,Brangsch J,Kader A,Saatz J,Mangarova DB,Zacharias M,Kempf WE,Schwaar T,Ponader M,Adams LC,Möckel J,Botnar RM,Taupitz M,Mägdefessel L,Traub H,Hamm B,Weller MG,Makowski MR

Published figure using ADAMTS4 polyclonal antibody (Product # PA1-1749A) in Immunohistochemistry

Mon May 23 00:00:00 EDT 2022

Versican is differentially regulated in the adventitial and medial layers of human vein grafts.

PloS one

Kenagy RD,Kikuchi S,Evanko SP,Ruiter MS,Piola M,Longchamp A,Pesce M,Soncini M,Deglise S,Fiore GB,Haefliger JA,Schmidt TA,Majesky MW,Sobel M,Wight TN

PA1-1749A was used in Immunohistochemistry to examine the production and distribution of versican and hyaluronan in intact human vein rings cultured ex vivo, veins perfused ex vivo, and cultured venous adventitial and smooth muscle cells.

Wed Mar 06 00:00:00 EST 2019

ADAMTS-4 and biglycan are expressed at high levels and co-localize to podosomes during endothelial cell tubulogenesis in vitro.

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Obika M,Vernon RB,Gooden MD,Braun KR,Chan CK,Wight TN

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

Wed Jan 01 00:00:00 EST 2014

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.

The international journal of biochemistry & cell biology

Ashlin TG,Buckley ML,Salter RC,Johnson JL,Kwan AP,Ramji DP

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

Wed Jan 01 00:00:00 EST 2014

Tumor necrosis factor-α induces ADAMTS-4 expression in human osteoarthritis chondrocytes.

Molecular medicine reports

Xue J,Wang J,Liu Q,Luo A

PA1-1749A was used in Western Blotting to demonstrate evidence of crosstalk between tumor necrosis factor-α and disintegrin and metalloproteinase with thrombospondin motifs 4 in relation to osteoarthritis cartilage degradation.

Sun Dec 01 00:00:00 EST 2013

Regulated proteolytic processing of Reelin through interplay of tissue plasminogen activator (tPA), ADAMTS-4, ADAMTS-5, and their modulators.

PloS one

Krstic D,Rodriguez M,Knuesel I

PA1-1749A was used in immunohistochemistry and western blot to study the role of tPA, ADAMTS-4, ADAMTS-5, and endogenous proteinase inhibitors in modulating the proteolytic cleavage of Reelin

Tue Apr 09 00:00:00 EDT 2013

Regulated proteolytic processing of Reelin through interplay of tissue plasminogen activator (tPA), ADAMTS-4, ADAMTS-5, and their modulators.

PloS one

Krstic D,Rodriguez M,Knuesel I

PA1-1749A was used in immunohistochemistry and western blot to study the role of tPA, ADAMTS-4, ADAMTS-5, and endogenous proteinase inhibitors in modulating the proteolytic cleavage of Reelin

Tue Apr 09 00:00:00 EDT 2013

The expression of a disintegrin and metalloproteinase with thrombospondin motifs 4 in human macrophages is inhibited by the anti-atherogenic cytokine transforming growth factor-β and requires Smads, p38 mitogen-activated protein kinase and c-Jun.

The international journal of biochemistry & cell biology

Salter RC,Arnaoutakis K,Michael DR,Singh NN,Ashlin TG,Buckley ML,Kwan AP,Ramji DP

Published figure using ADAMTS4 polyclonal antibody (Product # PA1-1749A) in Western Blot

Sun May 01 00:00:00 EDT 2011

Bioinformatics

Protein Aliases: a disintegrin and metalloproteinase; A disintegrin and metalloproteinase with thrombospondin motifs 4; a disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif, 4; ADAM; ADAM metallopeptidase with thrombospondin type 1 motif, 4; ADAM-TS 4; ADAM-TS4; ADAMs; ADAMTS family member; ADAMTS-4; ADMP-1; Aggrecanase 1; Aggrecanase-1; metalloendopeptidases; metalloproteinase

Gene Aliases: 3830423K05; ADAM-TS4; ADAMTS-2; ADAMTS-4; ADAMTS4; ADMP-1; KIAA0688; mKIAA0688; UNQ769/PRO1563

UniProt ID: (Bovine) Q9TT93, (Human) O75173, (Mouse) Q8BNJ2

Entrez Gene ID: (Bovine) 286806, (Human) 9507, (Pig) 100157276, (Mouse) 240913

Molecular Function: metalloprotease