Human ADAMTS4 MAb (Clone 416610)

Catalog #: MAB4307 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB4307
MAB4307-SP

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Human ADAMTS4 MAb (Clone 416610) Summary

Species Reactivity

Human

Specificity

Detects human ADAMTS4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) ADAMTS1, rhADAMTS5, rhADAMTS13, or rhADAMTSL1.2 is observed.

Source

Monoclonal Mouse IgG_2A Clone # 416610

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant human ADAMTS4
Phe213-Cys685
Accession # O75173

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

Recombinant Human ADAMTS4 (Catalog # 4307-AD) under non-reducing conditions only

Immunoprecipitation

25 µg/mL

Conditioned cell culture medium spiked with Recombinant Human ADAMTS4 (Catalog # 4307-AD), see our available Western blot detection antibodies

Immunocytochemistry

8-25 µg/mL

See below

Neutralization

Measured by its ability to neutralize Recombinant Human ADAMTS4 (5 µg/mL, Catalog # 4307-AD) cleavage of the substrate Recombinant Human Aggrecan G1-IGD-G2 Domain (100 µg/mL, Catalog # 1220-PG). The Neutralization Dose (ND₅₀) is typically 25 µg/mL.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunocytochemistry

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ADAMTS4 in SH‑SY5Y Human Cell Line. ADAMTS4 was detected in immersion fixed SH-SY5Y human neuroblastoma cell line using Mouse Anti-Human ADAMTS4 Monoclonal Antibody (Catalog # MAB4307) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: ADAMTS4

ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs 4), also known as aggrecanase-1, is a member of the family of secreted zinc proteases with a multi-domain structure (1‑3). The protein precursors consist of a signal peptide and the following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine-rich, and spacer. It is the only ADAMTS identified that has one TS type I motif. It is an active protease effectively cleaving alpha -2-macroglobulin and aggrecan at multiple sites, and is inhibited by TIMP-3 with inhibition constants in subnanomolar range (4‑6). It receives great attention due to the elevation in its mRNA level after treatment with Interleukin-1 (7). However, in a mouse model of osteoarthritis, ADAMTS4 knock-out mice did not exhibit any significant protective effect (8). ADAMTS4 consists of a signal peptide (aa 1 to 51), a pro domain (aa 52 to 212) and a mature chain (aa 213 to 837) containing the following domains: catalytic (aa 218 to 425), disintegrin (aa 437 to 519), TSP type-1 (aa 520 to 575), Cys-rich (aa 577 to 685) and spacer (aa 686 to 837). The amino acid sequence of rhADAMTS4 is 100%, 97% and 94% identical to that of chimpanzee, dog, and mouse/rat/bovine.

References

Totorella, M. D. et al. (1999) Science 284:1664.
Porter, S. et al. (2005) Biochem. J. 386:15.
Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
Tortorella, M. D. et al. (2004) J. Biol. Chem. 279:17554.
Struglics, A. et al. (2006) Osteoarthritis Cartilage. 14:101.
Kashiwagi, M. et al. (2001) J. Biol. Chem. 276:12501.
Pratta, M. A. et al. (2003) Arthritis Reum. 48:119.
Glasson, S. S. et al. (2004) Arthritis Reum. 50:2547.

Long Name

A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 4

Entrez Gene IDs

9507 (Human); 66015 (Rat)

Alternate Names

a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 4; ADAM metallopeptidase with thrombospondin type 1 motif, 4; ADAM-TS 4; ADAMTS-2; ADAMTS4; ADAM-TS4; ADAMTS-4; ADMP-1; ADMP-1EC 3.4.24.82; Aggrecanase 1; aggrecanase-1; EC 3.4.24; KIAA0688A disintegrin and metalloproteinase with thrombospondin motifs 4