Invitrogen

ADAMTS4 Polyclonal Antibody

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ADAMTS4 Antibody — Published figure using ADAMTS4 polyclonal antibody (Product # PA1-1749A)

ADAMTS4 Antibody in Immunofluorescence (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).

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

2 µg/mL

Immunofluorescence (IF)

2 µg/mL

Western Blot (WB)

1-3 µg/mL

Published Applications

Western Blot (WB)

See 4 publications below

Immunohistochemistry (Paraffin) (IHC (P))

See 1 publication below

Immunofluorescence (IF)

See 1 publication below

Immunohistochemistry (IHC)

See 1 publication below

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.

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

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's and other neurodegenerative diseases.

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

Bioinformatics

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

Gene Aliases: 3830423K05; ADAM-TS4; ADAMTS-2; ADAMTS-4; ADAMTS4; ADMP-1; BOS_2624; KIAA0688; mKIAA0688; UNQ769/PRO1563

UniProt ID: (Bovine) Q9TT93, (Human) Q5VTW2, (Mouse) Q8BNJ2

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

Molecular Function: enzyme modulator extracellular matrix glycoprotein extracellular matrix protein hydrolase metalloprotease protease protease inhibitor serine protease inhibitor