Human Fibronectin ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Fibronectin (FN) also known as LETS, is identified on the surfFN of fibroblasts by labeling with radioactive compounds or specific antibodies. Fibronectin is a 430,000-dalton dimeric glycoprotein that exists in 2 forms, termed cellular and plasma fibronectin. Cellular and plasma fibronectins are heterodimers consisting of similar but not identical polypeptides. These two forms of FN differ in biologic activity. Fibronectins bind cell surfFNs and various compounds including collagen, fibrin, heparin, DNA, and actin. Because fibronectin stimulates endocytosis in several systems and promotes the clearance of particulate material from the circulation, it could function in the clearance of C1q-coated material such as immune complexes or cellular debris. Fibronectins are involved in cell adhesion, cell motility, opsonization, would healing, and maintenance of cell shape. LETS, encoded on chromosome 8, is responsible for the LETS protein expression in humans. Because LETS has been implicated in tumorigenicity and cellular transformation, it is of interest that rearrangement or modifications in the number of chromosome 8 have been associated with certain forms of cancer. The standard used in this kit is isolated from human plasma with the molecular mass of 200-250KDa.

Plasma FN (soluble dimeric form) is secreted by hepatocytes. Cellular FN (dimeric or cross-linked multimeric forms), made by fibroblasts, epithelial and other cell types, is deposited as fibrils in the extracellular matrix. Ugl-Y1, Ugl-Y2 and Ugl-Y3 are found in urine.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 1.23 2.35 5.53 1.54 2.39 5.78 Standard deviation 0.063 0.099 0.265 0.12 0.177 0.399 CV (%) 5.1% 4.2% 4.8% 7.8% 7.4% 6.9% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 1.23 2.35 5.53 1.54 2.39 5.78 Standard deviation 0.063 0.099 0.265 0.12 0.177 0.399 CV (%) 5.1% 4.2% 4.8% 7.8% 7.4% 6.9% 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) 1.23 2.35 5.53 1.54 2.39 5.78 Mean (pg/ml) 1.23 2.35 5.53 1.54 2.39 5.78 Standard deviation 0.063 0.099 0.265 0.12 0.177 0.399 Standard deviation 0.063 0.099 0.265 0.12 0.177 0.399 CV (%) 5.1% 4.2% 4.8% 7.8% 7.4% 6.9% CV (%) 5.1% 4.2% 4.8% 7.8% 7.4% 6.9%

Fibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin. Fibronectins are involved in cell adhesion, cell motility, opsonization, wound healing, and maintenance of cell shape. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization. Participates in the regulation of type I collagen deposition by osteoblasts.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0439-CA Rabbit anti- human FN1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0439-DA Biotinylated goat anti- human FN1 polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0439-ST Lyophilized recombinant human FN1 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0439-CA Rabbit anti- human FN1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0439-DA Biotinylated goat anti- human FN1 polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0439-ST Lyophilized recombinant human FN1 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0439-CA Rabbit anti- human FN1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0439-CA Rabbit anti- human FN1 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0439-DA Biotinylated goat anti- human FN1 polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) EZ0439-DA Biotinylated goat anti- human FN1 polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0439-ST Lyophilized recombinant human FN1 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0439-ST Lyophilized recombinant human FN1 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

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) 0 156 312 625 1250 2500 5000 10000 O.D. 0.052 0.091 0.186 0.317 0.490 0.890 1.385 1.990 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.052 0.091 0.186 0.317 0.490 0.890 1.385 1.990 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.052 0.091 0.186 0.317 0.490 0.890 1.385 1.990 O.D. 0.052 0.091 0.186 0.317 0.490 0.890 1.385 1.990

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)