Human Tryptase/TPSAB1, B2 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Tryptases are serine proteases implicated in asthma and are highly expressed in human mast cells. They are derived from at least 4 nonallelic genes clustered on chromosome 16p13.3: TPSAB1, which represents the alpha and beta-I tryptase alleles; TPSB2, which represents the beta-II and beta-III tryptase alleles; TPSG1; and TPSD1. Elevated levels of serum tryptase occur in both anaphylactic and anaphylactoid reactions, but a negative test does not exclude anaphylaxis. Tryptase is less likely to be elevated in food allergy reactions as opposed to other causes of anaphylaxis.

TPSAB1: Isoform 1 and isoform 2 are expressed in lung, stomach, spleen, heart and skin; in these tissues, isoform 1 ispredominant. Isoform 2 is expressed in aorta, spleen, and breasttumor, with highest levels in the endothelial cells of some bloodvessels surrounding the aorta, as well as those surrounding thetumor and low levels, if any, in mast cells (at protein level) .

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 895 2837 6370 1384 3015 8572 Standard deviation 44.75 175.89 293.02 78.89 211.05 582.9 CV (%) 5% 6.2% 4.6% 5.7% 7% 6.8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 895 2837 6370 1384 3015 8572 Standard deviation 44.75 175.89 293.02 78.89 211.05 582.9 CV (%) 5% 6.2% 4.6% 5.7% 7% 6.8% 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) 895 2837 6370 1384 3015 8572 Mean (pg/ml) 895 2837 6370 1384 3015 8572 Standard deviation 44.75 175.89 293.02 78.89 211.05 582.9 Standard deviation 44.75 175.89 293.02 78.89 211.05 582.9 CV (%) 5% 6.2% 4.6% 5.7% 7% 6.8% CV (%) 5% 6.2% 4.6% 5.7% 7% 6.8%

Tryptase is the major neutral protease present in mast cells and is secreted upon the coupled activation-degranulation response of this cell type. May play a role in innate immunity. .

Catalog number Description Quantity Storage of opened/reconstituted material EZ0898-CA Rabbit anti- human Tryptase/TPSAB1, B2 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. EZ0898-DA Biotinylated goat anti- human Tryptase/TPSAB1, B2 polyclonal antibody (Detection Antibody) 500 μl, 12 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0898-ST Lyophilized recombinant human Tryptase/TPSAB1, B2 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 EZ0898-CA Rabbit anti- human Tryptase/TPSAB1, B2 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. EZ0898-DA Biotinylated goat anti- human Tryptase/TPSAB1, B2 polyclonal antibody (Detection Antibody) 500 μl, 12 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0898-ST Lyophilized recombinant human Tryptase/TPSAB1, B2 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 EZ0898-CA Rabbit anti- human Tryptase/TPSAB1, B2 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. EZ0898-CA Rabbit anti- human Tryptase/TPSAB1, B2 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. EZ0898-DA Biotinylated goat anti- human Tryptase/TPSAB1, B2 polyclonal antibody (Detection Antibody) 500 μl, 12 μg/mL (recommended dilution 1:100) EZ0898-DA Biotinylated goat anti- human Tryptase/TPSAB1, B2 polyclonal antibody (Detection Antibody) 500 μl, 12 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) EK0898-ST Lyophilized recombinant human Tryptase/TPSAB1, B2 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. EK0898-ST Lyophilized recombinant human Tryptase/TPSAB1, B2 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. 0.068 0.113 0.212 0.301 0.459 0.714 1.236 1.861 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.068 0.113 0.212 0.301 0.459 0.714 1.236 1.861 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.068 0.113 0.212 0.301 0.459 0.714 1.236 1.861 O.D. 0.068 0.113 0.212 0.301 0.459 0.714 1.236 1.861

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)