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Rat CX3CL1/Fractalkine

Rat CX3CL1/Fractalkine DuoSet ELISA Summary

Assay   Type

Solid   Phase Sandwich ELISA

Format

96-well   strip plate

Assay   Length

4.5   hours

Sample   Type & Volume Required Per Well

100 μL

Sensitivity

0.029   ng/mL

Assay   Range

195.0 -   12,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin   Plasma)

Specificity

Natural   and recombinant rat CX3CL1/Fractalkine

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

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant rat CX3CL1/Fractalkine. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

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: CX3CL1/Fractalkine

CX3CL1/Fractalkine is a transmembrane adhesion protein with a chemokine domain separated from the membrane by a mucin stalk. It is upregulated on many cell types during inflammation. The chemokine and mucin regions can be shed as a soluble chemokine that signals through the receptor CX3CR1. During extravasation, membrane-bound CX3CL1 traps leukocytes, then is cleaved to allow diapedesis. Soluble CX3CL1 protects neurons from microglial neurotoxicity, recruits macrophages for wound healing, recruits osteoclast precursors for bone resorption, and contributes to pathogenesis in coronary artery disease.

Long   Name:

chemokine   (C-X3-C motif) ligand 1

Entrez   Gene IDs:

6376   (Human); 20312 (Mouse); 89808 (Rat); 102117496 (Cynomolgus Monkey)

Alternate   Names:

ABCD-3;   C3Xkine; chemokine (C-X3-C motif) ligand 1; CX3C membrane-anchored chemokine;   CX3CL1; CXC3; CXC3C; FKN; Fractalkine; Neurotactin; neurotactin); NTNsmall   inducible cytokine subfamily D (Cys-X3-Cys), member 1 (fractalkine;   NTTSmall-inducible cytokine D1; SCYD1C-X3-C motif chemokine 1; small   inducible cytokine subfamily D (Cys-X3-Cys), member-1

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.

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

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

3Add 50 μL of Assay Diluent to each well.

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

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

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

7Aspirate and wash 5 times.

8Add 100 μL Substrate Solution to each well.

9Add 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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