Soluble Cluster of Differentiation 14, BioAssay™ ELISA Kit (Human) (sCD14, CD14 Antigen, Monocyte Differentiation Antigen CD14)

Kits and Assays / Kits-ELISA, BioAssay™

4°C

CD14 is a surface protein preferentially expressed on monocytes/macrophages. It binds lipopolysaccharide binding protein and recently has been shown to bind apoptotic cells. Alternative splicing results in multiple transcript variants encoding the same isoform.

Soluble Cluster of Differentiation 14, BioAssay™ ELISA Kit is a sandwich ELISA intended for the quantitative determination of sCD14 in human serum, plasma, cell culture supernates, tissue homogenates and urine.

Detection Range:
0.234-15ng/ml

Intra-assay Precision: %CV <8
Inter-assay Precision: %CV <10

Sensitivity:
The minimum detectable dose of human sCD14 is typically less than 0.229ng/ml.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined the mean O.D value of 20 replicates of the zero standard added by their three standard deviations.

Specificity:
This assay has high sensitivity and excellent specificity for detection of human sCD14. No significant crossreactivity or interference between human sCD14 and analogs was observed.
Note: Limited by current skills and knowledge, it is impossible for us to complete the crossreactivity detection between human sCD14 and all the analogs. Therefore, crossreaction may still exist.

Kit Components:
227920A: Microtiter Strips, 1x48 wells or 1x96 wells. Pre-coated; ready to use.
227920B: Standard, 2x1vial
227920C: sCD14 (Biotin), 100X, 1x60ul or 1x120ul
227920D: Avidin (HRP), 100X, 1x60ul or 1x120ul
227920E: Biotin Diluent, 1x7.5ml or 1x15ml
227920F: Avidin (HRP) Diluent, 1x7.5ml or 1x15ml
227920G: Sample Diluent, 1x25ml or 1x50ml
227920H: Wash Buffer, 25X, 1x10ml or 1x20ml
227920J: TMB Substrate, 1x5ml or 1x10ml
227920K: Stop Solution, 1x5ml or 1x10ml

Note: The Stop Solution (227920K) provided with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Storage and Stability:
Store kit at 4°C. The unopended kit is stable for 6 months after receipt. Once opened, the kit is stable up to 4 weeks at 4°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Assay Summary:
1. Prepare reagents, standards and samples as instructed.
2. Add 100ul standard or sample to each well. Incubate for 2 hours at 37°C
3. Remove the liquid of each well do not wash.
4. Add 100ul sCD14 (Biotin), 1X to each well. Incubate for 1 hour at 37°C
5. Aspirate and wash 3 times.
6. Add 100ul Avidin (HRP), 1X to each well. Incubate for 1 hour at 37°C
7. Aspirate and wash 5 times.
8. Add 90ul TMB substrate to each well Incubate for 15-30 minutes at 37°C. Protect from light
9. Add 50ul Stop Solution to each well. Read at 450nm within 5 minutes.