Human CD14 ELISA Kit

Human CD14 ELISA Kit Features

Product Name            Human CD14 ELISA Kit
Assay Type                   Solid Phase Sandwich ELISA
Format                           96-well strip plate
Assay Length              4.5 hours
Sample Volume         Serum ,Plasma, cell culture media , 100ul
Detection Method    Colorimetric
Sensitivity                   4 pg/mL
Assay Range                8.2-6000 pg/mL
Reactivity                     Human
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               CD14
Gene ID                         929
Swissprot ID               P08571

Recovery

Linearity

Standard Curve: Human CD14 ELISA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Standards
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µl of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Wash and add 100 µl of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Wash and add 100 µl of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µl of Stop Solution to each well.
11. Read at 450 nm/570 nm.

Background

CD14 (cluster of differentiation 14), also known as CD14, is a human gene. The protein encoded by this gene is a component of the innate immune system. CD14 exists in two forms, one anchored to the membrane by a glycosylphosphatidylinositol tail (mCD14), the other a soluble form (sCD14). Soluble CD14 either appears after shedding of mCD14 (48 kDa) or is directly secreted from intracellular vesicles (56 kDa). CD14 is an acute phase glycoprotein that binds lipopolysaccharide endotoxins (LPS) from Gram-negative bacteria. It is primarily expressed on monocytes, macrophages, and neutrophils, and soluble forms can be secreted or shed from the cell surface. CD14 accepts LPS from its initial binding by LBP, and it can further transfer LPS to the TLR4/MD2 complex. LPS acctivation of TLR4 triggers inflammatory activation. CD14 also contributes to the response to TLR3 ligands, phosphoinositides, as well as the surfactant proteins SP-A and SP-D.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status 

Human CD14 ELISA Kit is for Research Use Only.

• Manual
• Assayprocedure
• MSDS