Overview
| Product Name |
Human ALCAM/Cd166 ELISA Kit PicoKine™
See all ALCAM/CD166 primary antibodies, ELISA kits and proteins |
| SKU/Catalog Number |
EK0995 |
| Storage & Handling |
Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.) |
| Description |
Detect Human Cd166/ALCAM with <10pg/ml sensitivity. Format: 96-well plate with removable strips. Compatible samples: cell culture supernates, cell lysates, serum and plasma (heparin, EDTA). This is a TMB colorimetric sandwich ELISA kit with short assay time and fast experiment set up. Cd166/ALCAM tissue specificity: Spleen, placenta, liver, and weakly in liver. Expressed by activated T-cells, B-cells, monocytes and thymic epithelial cells. Expressed by neurons in the brain. Restricted expression in tumor cell lines. Preferentially expressed in highly metastasizing melanoma cell lines. . |
| Cite This Product |
Human ALCAM/Cd166 ELISA Kit PicoKine™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0995)
|
Product Specs
| Reactivity/Species |
Human |
| Applications |
ELISA
*Our Boster Guarantee covers the use of this product in the above
tested applications.
**For positive and negative control design, consult "Tissue specificity" under
"Protein Target Info" tab. |
| Sample Type |
cell culture supernates, cell lysates, serum and plasma (heparin, EDTA). |
| Sensitivity |
<10pg/ml |
| Assay Range |
312pg/ml-20,000pg/ml |
| Immunogen/Standard |
Expression system for standard: NSO; Immunogen sequence: W28-A526 |
| Antibody Clonalities |
Capture antibody|Detection antibody
monoclonal antibody from mouse|polyclonal antibody from goat |
| Cross Reactivity |
There is no detectable cross-reactivity with other relevant proteins. |
| Pack Size |
96wells/kit, with removable strips. |
Gene/Protein Basic Information For ALCAM (Source: Uniprot.org, NCBI)
| Uniprot Id | Q13740 |
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| NCBI Gene Id | 214 |
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| Species Of This Entry | Human |
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| Gene Name | ALCAM |
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| Protein Name | CD166 antigen |
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| Alternative Names | ALCAM/CD166|activated leucocyte cell adhesion molecule; activated leukocyte cell adhesion moleculeFLJ38514; ALCAM; CD166 antigen; CD166; MEMDMGC71733 |
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| Molecular Weight | 65102 |
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| Post Tranlational Modifications | Phosphorylation; Glycosylation; Cleavage; Methylation; Biotinylation; Acetylation; Deglycosylation; Ubiquitination; Myristoylation |
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| Subcellular Localization | Cell membrane; Single-pass type I membrane protein. Cell projection, axon. Cell projection, dendrite. Detected at the immunological synapse, i.e, at the contact zone between antigen-presenting dendritic cells and T-cells (PubMed:15294938, PubMed:16352806). Colocalizes with CD6 and the TCR/CD3 complex at the immunological synapse (PubMed:15294938).; [Isoform 3]: Secreted. |
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| Gene Location | 3q13.11, on Chromasome 3, gene sequence: NC_000003.12 (105366713..105576900) |
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*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".
Ontology For ALCAM (Source: Uniprot.org, NCBI)
| Protein Function | Cell adhesion molecule that mediates both heterotypic cell-cell contacts via its interaction with CD6, as well as homotypic cell-cell contacts (PubMed:7760007, PubMed:15496415, PubMed:15048703, PubMed:16352806, PubMed:23169771, PubMed:24945728). Promotes T-cell activation and proliferation via its interactions with CD6 (PubMed:15048703, PubMed:16352806, PubMed:24945728). Contributes to the formation and maturation of the immunological synapse via its interactions with CD6 (PubMed:15294938, PubMed:16352806). Mediates homotypic interactions with cells that express ALCAM (PubMed:15496415, PubMed:16352806). Required for normal hematopoietic stem cell engraftment in the bone marrow (PubMed:24740813). Mediates attachment of dendritic cells onto endothelial cells via homotypic interaction (PubMed:23169771). Inhibits endothelial cell migration and promotes endothelial tube formation via homotypic interactions (PubMed:15496415, PubMed:23169771). Required for normal organization of the lymph vessel network. Required for normal hematopoietic stem cell engraftment in the bone marrow. Plays a role in hematopoiesis; required for normal numbers of hematopoietic stem cells in bone marrow. Promotes in vitro osteoblast proliferation and differentiation (By similarity). Promotes neurite extension, axon growth and axon guidance; axons grow preferentially on surfaces that contain ALCAM. Mediates outgrowth and pathfinding for retinal ganglion cell axons (By similarity). |
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| Tissue Specificity | Detected on hematopoietic stem cells derived from umbilical cord blood (PubMed:24740813). Detected on lymph vessel endothelial cells, skin and tonsil (PubMed:23169771). Detected on peripheral blood monocytes (PubMed:15048703). Detected on monocyte-derived dendritic cells (at protein level) (PubMed:16352806). Detected at low levels in spleen, placenta, liver (PubMed:9502422). Expressed by activated T-cells, B-cells, monocytes and thymic epithelial cells (PubMed:7760007). Isoform 1 and isoform 3 are detected in vein and artery endothelial cells, astrocytes, keratinocytes and artery smooth muscle cells (PubMed:15496415). Expressed by neurons in the brain. Restricted expression in tumor cell lines. Detected in highly metastasizing melanoma cell lines (PubMed:9502422). |
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| Interacting Proteins | CD6; LGALS1; LGALS3 |
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| Related Diseases | Tissue Adhesions; Neoplasms; Malignant Neoplasms; Neoplasm Metastasis; Carcinoma; Malignant Neoplasm Of Breast; Cell Invasion; Mammary Neoplasms; Melanoma; Malignant Paraganglionic Neoplasm; Neoplasm Invasiveness; Colorectal Cancer; Tumor Progression; Cell Transformation, Neoplastic; Colorectal Neoplasms; Nervousness; Malignant Neoplasm Of Prostate; Skin Neoplasms; Adenocarcinoma |
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| Related Pathways | Cell Adhesion; Cell Migration; Cell Proliferation; Localization; Cell-cell Adhesion; Immune Response; Cell Differentiation; Cell Growth; Angiogenesis; Cell Activation; T Cell Activation; Rna Interference; Cell Motility; Regeneration; Cell Cycle; Glycosylation; Pathogenesis; Tube Formation; Chemotaxis; Ovulation |
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| 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. |
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| Scientific References | PMID: 7760007 by Bowen M.A., et al. Cloning, mapping, and characterization of activated leukocyte-cell adhesion molecule (ALCAM), a CD6 ligand.
PMID: 15496415 by Ikeda K., et al. Molecular isolation and characterization of a soluble isoform of activated leukocyte cell adhesion molecule that modulates endothelial cell function. |
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Kit Components And Assay QC Data Details Of Human ALCAM/Cd166 ELISA Kit PicoKine™
*The quality control (QC) data in this section is obtained from Boster's internal QC results and is for reference only. It may differ from the users' lab test results. The users can expect to generate data with similar linearity and quality demonstrated in the typical data but may not achieve exactly the same O.D. values.
| Catalog number | Description | Quantity |
| EK0995-CAP | 96-well plate precoated with anti-Human ALCAM/CD166 antibody | 1 |
| EK0995-ST | lyophilized recombinant Human ALCAM/CD166 standard | 20ng/tube |
| EK0995-DA | biotinylated anti-Human ALCAM/CD166 antibody | 130ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex(ABC) | 130ul |
| 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 |
| PLA-SEA | Adhesive cover | 4 |
- 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; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
This data is generated from a recent batch of EK0995 Human ALCAM/Cd166 ELISA Kit PicoKine™. It is not necessarily the same data set used to generate the standard curve shown in the image of this product. TMB reaction incubate at 37°C for 15-25min
| Concentration(pg/ml) | 0 | 312 | 625 | 1250 | 2500 | 5000 | 10000 | 20000 |
| O.D. | 0.094 | 0.276 | 0.390 | 0.635 | 1.148 | 1.589 | 1.994 | 2.274 |
We measured random samples of Human ALCAM/Cd166 ELISA Kit PicoKine™ within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
| Intra-Assay Precision | Inter-Assay Precision |
|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean(pg/ml) | 648 | 1967 | 8283 | 688 | 2156 | 8924 |
| Standard deviation | 37.58 | 145.55 | 563.24 | 48.16 | 198.35 | 651.45 |
| CV(%) | 5.8% | 7.4% | 6.8% | 7% | 9.2% | 7.3% |
We measured the performance consistency of Human ALCAM/Cd166 ELISA Kit PicoKine™ to ensure the reproducibility of the assays. Three samples with differing target protein concentrations were assayed using four different lots.
Reproducibility
To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.
| Lots | Lot1 (pg/ml) | Lot2 (pg/ml) | Lot3 (pg/ml) | Lot4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|
| Sample 1 | 648 | 630 | 693 | 657 | 657 | 22.94 | 3.4% |
| Sample 2 | 1967 | 2006 | 1987 | 2137 | 2024 | 66.54 | 3.2% |
| Sample 3 | 8283 | 8482 | 8134 | 7570 | 8117 | 339.21 | 4.1% |
*number of samples for each test n=16.
Specific Protocols
Boster provides comprehensive technical information for WB, IHC/IF/ICC, Flow Cytometry sample preparation protocols, assay protocols, troubleshooting tips and assay optimization tips.
See all ALCAM/CD166 primary antibodies,
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