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Mouse/Rat TNF RI/TNFRSF1A

Mouse/Rat TNF RI/TNFRSF1A Quantikine ELISA Kit

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), Serum (10 uL)

Sensitivity

2.4   pg/mL

Assay   Range

15.6 -   1,000 pg/mL (Cell Culture Supernates, Serum)

Specificity

Natural   and recombinant mouse and rat TNF RI

 

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 Mouse/Rat TNF RI immunoassay is a 4.5 hour solid-phase ELISA designed to measure TNF RI in mouse or rat cell culture supernates and serum. It contains E. coli-expressed recombinant mouse TNF RI and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse TNF RI. Results obtained using natural mouse or rat TNF RI showed dose response 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 TNF RI.

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: TNF RI/TNFRSF1A

TNF RI (TNF receptor 1; TNF R-p55/p60) is a transmembrane receptor for TNF-alpha and Lymphotoxin-alpha/TNF-beta. A soluble decoy form of the receptor is shed by ADAM-17/TACE in response to inflammatory stimulation. TNF RI signaling through NFkB is essential for the development of lymph node germinal centers and Peyer’s patches and for combating intracellular pathogens such as Listeria.

Long   Name:

Tumor   Necrosis Factor Receptor I

Entrez   Gene IDs:

7132   (Human); 21937 (Mouse); 25625 (Rat); 102135285 (Cynomolgus Monkey)

Alternate   Names:

CD120a   antigen; CD120a; FPF; p55; p55-R; p60; TNF RI; TNFARMGC19588; TNF-R; TNF-R1;   TNFR1TBP1; TNFR55; TNF-R55; TNFR60; TNFRI; TNF-RI; TNF-R-I; TNFR-I; TNFRSF1A;   tumor necrosis factor binding protein 1; Tumor necrosis factor receptor 1;   tumor necrosis factor receptor 1A isoform beta; tumor necrosis factor   receptor superfamily member 1A; tumor necrosis factor receptor superfamily,   member 1A; tumor necrosis factor receptor type 1; Tumor necrosis factor   receptor type I; tumor necrosis factor-alpha receptor

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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