Rat Matrilin-4 ELISA Kit PicoKine®

Matrilin-4 is a 73 kDa secreted glycoprotein that is a member of the matrilin family of the von Willebrand Factor-A (vWA) domain-containing superfamily. This gene is mapped to 3q42. Matrilins are modular extracellular matrix proteins that serve as adaptors and linkers for other matrix proteins. Matrilin-4, like Matrilin?2, has a broad distribution in both cartilage and in loose connective tissue such as dermis, lung and kidney, while Matrilins 1 and 3 are limited to cartilage. Matrilin-4 is present in nervous tissue and is abundant in the brain. Mature mouse Matrilin-4 shares 98%, 90%, 89% and 66% amino acid (aa) identity with rat, human, canine and chicken Matrilin-4, respectively. The 624 aa mouse Matrilin-4 contains a 22 aa signal sequence, two potential glycosylation sites, and four cysteine-rich EGF-like domains placed between two vWA domains. A short isoform lacks the N-terminal vWA domain (aa 28?217) . A C-terminal alpha -helix/coiled-coil region (aa 590?623) by which multimers are formed is often proteolytically removed so that Matrilin-4 is found as a mixture of monomers with homo- or hetero- dimers and trimers. Matrilin-4 forms multimers with Matrilins 1 and 2 but not with Matrilin-3. The N-terminal vWA domains of Matrilins associate with collagen IV microfibrils via the proteoglycans biglycan and decorin, linking the fibrils with other matrix constituents aggrecan and collagen II. Matrilins also show calcium-dependent binding to the cartilage oligomeric matrix protein (COMP) ; this interaction is of high affinity for oligomeric Matrilin-4 and somewhat lower affinity for monomeric Matrilin-4. Functions and distributions of Matrilins overlap enough so that knockouts of Matrilins 1, 2 and 3 lack obvious phenotypes.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 37 37 43 38 38 2.48 6.5% Sample 2 157 186 160 160 165 11.75 7.1% Sample 3 741 797 765 814 779 28.23 3.6% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 37 37 43 38 38 2.48 6.5% Sample 1 37 37 43 38 38 2.48 6.5% Sample 2 157 186 160 160 165 11.75 7.1% Sample 2 157 186 160 160 165 11.75 7.1% Sample 3 741 797 765 814 779 28.23 3.6% Sample 3 741 797 765 814 779 28.23 3.6%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 37 157 741 43 186 939 Standard deviation 1.92 9.89 35.57 2.11 13.76 49.77 CV (%) 5.2% 6.3% 4.8% 4.9% 7.4% 5.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 37 157 741 43 186 939 Standard deviation 1.92 9.89 35.57 2.11 13.76 49.77 CV (%) 5.2% 6.3% 4.8% 4.9% 7.4% 5.3% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 37 157 741 43 186 939 Mean (pg/ml) 37 157 741 43 186 939 Standard deviation 1.92 9.89 35.57 2.11 13.76 49.77 Standard deviation 1.92 9.89 35.57 2.11 13.76 49.77 CV (%) 5.2% 6.3% 4.8% 4.9% 7.4% 5.3% CV (%) 5.2% 6.3% 4.8% 4.9% 7.4% 5.3%

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.081 0.18 0.254 0.402 0.634 1.116 1.931 3.045 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.081 0.18 0.254 0.402 0.634 1.116 1.931 3.045 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.081 0.18 0.254 0.402 0.634 1.116 1.931 3.045 O.D. 0.081 0.18 0.254 0.402 0.634 1.116 1.931 3.045

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.