CD14 ELISA Kit

Catalog No. ABIN1672772

Product Details CD14 ELISA Kit

Target: CD14

Binding Specificity: AA 18-345

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156-10000 pg/mL

Minimum Detection Limit: 156 pg/mL

Application: ELISA

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse CD14

Brand: PicoKine™

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: A18-P345

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: NSO
Immunogen sequence: A18-P345

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Plate: Pre-coated

Protocol: mouse CD14 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for CD14 has been precoated onto 96-well plates. Standards(NSO, A18-P345) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD14 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse CD14 amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 10000pg/mL, 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312pg/mL, 156pg/mL mouse CD14 standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse CD14 standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(ng/ml): 0.98, Standard deviation: 0.038, CV(%): 3.9
• Sample 2: n=16, Mean(ng/ml): 3.11, Standard deviation: 0.162, CV(%): 5.2
• Sample 3: n=16, Mean(ng/ml): 6.23, Standard deviation: 0.355, CV(%): 5.7,
• Sample 1: n=24, Mean(ng/ml): 1.03, Standard deviation: 0.081, CV(%): 7.9
• Sample 2: n=24, Mean(ng/ml): 3.47, Standard deviation: 0.249, CV(%): 7.2
• Sample 3: n=24, Mean(ng/ml): 6.72, Standard deviation: 0.423, CV(%): 6.3

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References for CD14 ELISA Kit (ABIN1672772)

Jiang, Chen, Xie, Ji, Liu, Qiu, Zhu, Wang: "Intrapulmonary expression of scavenger receptor and CD14 and their relation to local inflammatory responses to endotoxemia in mice." in: World journal of surgery, Vol. 27, Issue 1, pp. 1-9, (2003) (PubMed).

Target Details for CD14

Target: CD14

Alternative Name: CD14 (CD14 Products)

Synonyms: CD14 molecule ELISA Kit, CD14 antigen ELISA Kit, CD14 ELISA Kit, Cd14 ELISA Kit

Background:

Protein Function: In concert with LBP, binds to monomeric lipopolysaccharide and delivers it to the MD-2/TLR4 complex, thereby mediating the innate immune response to bacterial lipopolysaccharide (LPS). Acts via MyD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Up-regulates cell surface molecules, including adhesion molecules (By similarity). .

Background: CD14, Cluster of differentiation 14, ingle-copy gene encoding 2 protein forms: a 50- to 55-kD glycosylphosphatidylinositol-anchored membrane protein(mCD14) and a monocyte or liver-derived soluble serum protein(sCD14) that lacks the anchor. By in situ hybridization and study of somatic cell hybrid DNA that the gene is located at bands 5q23-q31. CD14 acts as a co-receptor(along with the Toll-like receptor TLR 4 and MD-2) for the detection of bacterial lipopolysaccharide(LPS). CD14 can bind LPS only in the presence of lipopolysaccharide-binding protein(LBP). Although LPS is considered its main ligand, CD14 also recognizes other pathogen-associated molecular patterns.

Synonyms: Monocyte differentiation antigen CD14,Myeloid cell-specific leucine-rich glycoprotein,CD14,Cd14,

Full Gene Name: Monocyte differentiation antigen CD14

Cellular Localisation: Cell membrane, Lipid-anchor, GPI-anchor.

Gene ID: 12475

UniProt: P10810

Pathways: TLR Signaling, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Toll-Like Receptors Cascades