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Human TRAIL/TNFSF10

Human TRAIL/TNFSF10 Quantikine ELISA Kit Summary

Assay   Type

Solid   Phase Sandwich ELISA

Format

96-well   strip plate

Assay   Length

4.5   hours

Sample   Type & Volume Required Per Well

Cell   Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (50 uL), EDTA Plasma   (50 uL), Heparin Plasma (50 uL), Saliva (25 uL)

Sensitivity

7.87   pg/mL

Assay   Range

15.6 -   1,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma,   Heparin Plasma, Saliva)

Specificity

Natural   and recombinant human TRAIL

 

Cross-reactivity

<   0.5% cross-reactivity observed with available related molecules.< 50% cross-species   reactivity observed with species tested

Interference

No   significant interference observed with available related molecules.

Product Summary

The Quantikine Human TRAIL Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human TRAIL in cell culture supernates, serum, plasma, cell lysates, and saliva. It contains NS0-expressed recombinant human TRAIL and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human TRAIL showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human TRAIL.

Preparation and Storage

Shipping

The   product is shipped at ambient temperature. Upon receipt, store it immediately   at the temperature recommended below.

Storage

Store   the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TRAIL/TNFSF10

TNF-related apoptosis-inducing ligand (TRAIL), also known as Apo-2 ligand and TNFSF10, is a type II transmembrane protein with a carboxy-terminal extracellular domain that exhibits homology to other TNF superfamily members. Among TNF superfamily members, TRAIL is the most homologous to Fas Ligand, sharing 28% amino acid sequence identity in their extracellular domains. Human TRAIL shares 65% amino acid sequence identity with mouse TRAIL and is active on mouse cells.

Long   Name:

TNF-related   Apoptosis-inducing Ligand

Entrez   Gene IDs:

8743   (Human); 22035 (Mouse)

Alternate   Names:

Apo-2   ligand; APO2L; Apo-2Ltumor necrosis factor (ligand) family, member 10;   APO2Ltumor necrosis factor apoptosis-inducing ligand splice variant delta;   CD253 antigen; CD253; Protein TRAIL; TL2; TNF-related apoptosis-inducing   ligand; TNFSF10; TRAIL; TRAILTNF-related apoptosis inducing ligand TRAIL;   tumor necrosis factor (ligand) superfamily, member 10; tumor necrosis factor   ligand superfamily member 10

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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