Mouse ALCAM PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of mouse ALCAM 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 ALCAM
Protein name: CD166 antigen
Gene Full Name: activated leukocyte cell adhesion molecule
abbreviation: ALCAM
Background: CD166 antigen is a 100-105 kD typeI transmembrane glycoprotein that is a member of the immunoglobulin superfamily of proteins. In humans it is encoded by the ALCAM gene. The gene was mapped to human chromosome 3q13.1-q13.2 by fluorescence in situ hybridization of cDNA probes to metaphase chromosomes. It is also called CD166 (cluster of differentiation 166), MEMD, SC-1/DM-GRASP/BEN in the chicken, and KG-CAM in the rat. It is expressed on activated T cells, activated monocytes, epithelial cells, fibroblasts, neurons, melanoma cells, and also in sweat and sebaceous glands. CD166 protein expression is reported to be upregulated in a cell line deriving from a metastasizing melanoma. CD166 plays an important role in mediating adhesion interactions between thymic epithelial cells and CD6+ cells during intrathymic T cell development. Recently, CD166 has also been used as a potential cancer stem cell marker.
ID:Q61490
ID:11658

alias

ALCAM protein | CD166 antigen | Protein MEMD | Q13740

Origin

Nativ and recombinant mouse ALCAM

Antibodies

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

Immunogene

NSO, W28-K527

sensitivity

<10pg/ml

detection range

78pg/ml-5000pg/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(pg/ml)|121|495|1946|93.2|513|2209
Standard deviation|5.93|20.79|103.13|6.43|31.29|88.36
CV(%)|4.9|4.2|5.3|6.9|6.1|4.0

TMB(tetramethylbenzidine) incubation time

25-30

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 ALCAM ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for ALCAM has been precoated onto 96-well plates. Standards(NSO, W28-K527) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for ALCAM 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 ALCAM amount of sample captured in plate.

Assay Procedure Step 1

Aliquot 0.1ml per well of the 5000pg/ml, 2500pg/ml, 1250pg/ml, 625pg/ml, 313pg/ml, 156pg/ml, 78pg/mll mouse ALCAM 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 ALCAM standard solution and each sample be measured in duplicate.

tipical result

Concentration(pg/ml)|0|78|156|312|625|1250|2500|5000 O.D.|0.066|0.098|0.164|0.294|0.544|1.007|1.821|2.388

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- mouse ALCAM antibody|1
ST0000-10|Lyophilized recombinant mouse ALCAM standard|10ng/tube�2
AR1107|Biotinylated anti- mouse ALCAM antibody|130µl(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex (ABC)|130µl(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