Human Angiogenin/ANG ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Angiogenin (Ang) also known as ribonuclease 5 is a protein that in humans is encoded by the ANG gene. Angiogenin is a potent stimulator of new blood vessel formation. It hydrolyzes cellular tRNAs resulting in decreased protein synthesis and is similar to pancreatic ribonuclease. Hooper et al. (2003) reviewed the evidence that angiogenins are involved in host defense and noted that inflammation provokes upregulated ANG mRNA expression in liver and an increase in detectable ANG protein in serum. Weremowicz et al. (1989, 1990) assigned the human angiogenin gene to chromosome 14q11.

Actin Binding Proteins, Actin, etc., Angiogenesis, Cancer, Cancer Metabolism, Cardiovascular, Cytoskeleton, Cytoskeleton/ECM, DNA/RNA, Epigenetics and Nuclear Signaling, Hypoxia, Invasion/Microenvironment, Metabolism, Metabolism Processes, Microfilaments, Neurodegenerative Disease, Neurology Process, Neuroscience, Pathways and Processes, Response To Hypoxia, RNA Processing, Signal Transduction, Translation

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 312 672 1663 635 1856 3489 Standard deviation 9.4 23.52 54.9 45.72 165.2 300 CV (%) 3% 3.5% 3.3% 7.2% 8.9% 8.6% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 312 672 1663 635 1856 3489 Standard deviation 9.4 23.52 54.9 45.72 165.2 300 CV (%) 3% 3.5% 3.3% 7.2% 8.9% 8.6% 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) 312 672 1663 635 1856 3489 Mean (pg/ml) 312 672 1663 635 1856 3489 Standard deviation 9.4 23.52 54.9 45.72 165.2 300 Standard deviation 9.4 23.52 54.9 45.72 165.2 300 CV (%) 3% 3.5% 3.3% 7.2% 8.9% 8.6% CV (%) 3% 3.5% 3.3% 7.2% 8.9% 8.6%

Binds to actin on the surface of endothelial cells; once bound, angiogenin is endocytosed and translocated to the nucleus. Stimulates ribosomal RNA synthesis including that containing the initiation site sequences of 45S rRNA. Cleaves tRNA within anticodon loops to produce tRNA-derived stress-induced fragments (tiRNAs) which inhibit protein synthesis and triggers the assembly of stress granules (SGs) . Angiogenin induces vascularization of normal and malignant tissues. Angiogenic activity is regulated by interaction with RNH1 in vivo. .

Catalog number Description Quantity Storage of opened/reconstituted material EZ0305-CA Rabbit anti- human ANG 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. EZ0305-DA Biotinylated goat anti- human ANG polyclonal antibody (Detection Antibody) 500 μl, 8.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0305-ST Lyophilized recombinant human ANG 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 EZ0305-CA Rabbit anti- human ANG 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. EZ0305-DA Biotinylated goat anti- human ANG polyclonal antibody (Detection Antibody) 500 μl, 8.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0305-ST Lyophilized recombinant human ANG 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 EZ0305-CA Rabbit anti- human ANG 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. EZ0305-CA Rabbit anti- human ANG 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. EZ0305-DA Biotinylated goat anti- human ANG polyclonal antibody (Detection Antibody) 500 μl, 8.3 μg/mL (recommended dilution 1:100) EZ0305-DA Biotinylated goat anti- human ANG polyclonal antibody (Detection Antibody) 500 μl, 8.3 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) EK0305-ST Lyophilized recombinant human ANG 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. EK0305-ST Lyophilized recombinant human ANG 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.)

Nucleus . Secreted, extracellular space, extracellular matrix, basement membrane. Nucleus, nucleolus. Rapidly endocytosed by target cells and translocated to the nucleus where it accumulates in the nucleolus and binds to DNA.

Concentration (pg/ml) 0 78 156 313 625 1250 2500 5000 O.D. 0.047 0.116 0.169 0.265 0.506 0.927 1.534 2.375 Concentration (pg/ml) 0 78 156 313 625 1250 2500 5000 O.D. 0.047 0.116 0.169 0.265 0.506 0.927 1.534 2.375 Concentration (pg/ml) 0 78 156 313 625 1250 2500 5000 Concentration (pg/ml) 0 78 156 313 625 1250 2500 5000 O.D. 0.047 0.116 0.169 0.265 0.506 0.927 1.534 2.375 O.D. 0.047 0.116 0.169 0.265 0.506 0.927 1.534 2.375

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)