Mouse CD14 PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of mouse CD14 in cell culture supernates, cell lysates, tissue homogenates, serum and plasma (heparin, EDTA).

spezie

Mouse

Cross Reaction

There is no detectable cross-reactivity with other relevant proteins.

detection target

Mouse CD14
Protein name: Monocyte differentiation antigen CD14
Gene Full Name: CD14 molecule
abbreviation: CD14
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.
ID:P10810
ID:12475

alias

CD 14|CD14 antigen|CD14 molecule|Monocyte differentiation antigen CD14|Myeloid cell-specific leucine-rich glycoprotein

Origin

Nativ and recombinant mouse CD14

Antibodies

a monoclonal antibody from rat|a biotinylated polyclonal antibody from goat

Immunogene

NSO, A18-P345

sensitivity

<10pg/ml

detection range

156pg/ml-10000pg/ml

pack size

1x96 Wells

Reproducibility

|Intra-Assay Precision|Inter-Assay Precision
Sample|1|2|3|1|2|3
n|16|16|16|24|24|24
Mean(ng/ml)|0.98|3.11|6.23|1.03|3.47|6.72
Standard deviation|0.038|0.162|0.355|0.081|0.249|0.423
CV(%)|3.9|5.2|5.7|7.9|7.2|6.3

TMB(tetramethylbenzidine) incubation time

15-20

storage and Stability

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

Principle of the Elisa

Boster′s 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 Step 1

Aliquot 0.1ml 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 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See ﾓSample Dilution Guidelineﾔ above for details. It is recommended that each mouse CD14 standard solution and each sample be measured in duplicate.

tipical result

Concentration(pg/ml)|0|156|312|625|1250|2500|5000|10000 O.D.|0.064|0.077|0.129|0.214|0.396|0.697|1.142|2.029

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- mouse CD14 antibody|1
ST0000-10|Lyophilized recombinant mouse CD14 standard|10ng/tubex2
AR1107|Biotinylated anti- mouse CD14 antibody|130ul(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex(ABC)|130ul(dilution 1:100)
AR1106-1|Sample diluent buffer|30ml
AR1106-2|Antibody diluent buffer|12ml
AR1106-3|ABC diluent buffer|12ml
AR1104|TMB color developing agent|10ml
AR1105|TMB stop solution|10ml
-|Adhesive cover|4