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Rat ALCAM/CD166 EZ-Set™ ELISA Kit (DIY Antibody Pairs)

Rat ALCAM/CD166 ELISA Kit EZ-Set™ (DIY Antibody Pairs) (antibody pairs and standards for assay development) . Quantitate Rat Alcam in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) .

Product Specifications

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 human gene was mapped to 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. This gene is expressed on activated T cells, activated monocytes, epithelial cells, fibroblasts, neurons, melanoma cells, and also in sweat and sebaceous glands. 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.

Synonyms

CD166 antigen; Activated leukocyte cell adhesion molecule; HB2 ; KG-CAM ; Protein MEMD; SB-10 antigen ; CD166; Alcam

Gene Name

Alcam

Gene ID

79559

UniProt

O35112

Reactivity

Rat

Cross Reactivity

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

Immunogen

Expression system for standard: NS0; Immunogen sequence: W28-K527

Tissue Specificity

Strongest expression in the lung, then brain, liver, and kidney. Present in the somatosensory system, basal ganglia, cortex, olfactory system, and circumventricular organs.

Applications

ELISA

Field of Research

Adaptive Immunity, B Cells, Immunology, T Cells

Sample Type

Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) .

Detection Range

78 pg/mL - 5,000 pg/mL

Function

Cell adhesion molecule that mediates both heterotypic cell-cell contacts via its interaction with CD6, as well as homotypic cell-cell contacts. Promotes T-cell activation and proliferation via its interactions with CD6 (By similarity) . Contributes to the formation and maturation of the immunological synapse via its interactions with CD6 (By similarity) . Mediates homotypic interactions with cells that express ALCAM. Mediates attachment of dendritic cells onto endothelial cells via homotypic interaction. Inhibits endothelial cell migration and promotes endothelial tube formation via homotypic interactions. 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 (By similarity) . Mediates outgrowth and pathfinding for retinal ganglion cell axons (By similarity) . .

Components

Catalog number Description Quantity Storage of opened/reconstituted material EZ1572-CA Goat anti-rat ALCAM/CD166 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1572-DA Biotinylated goat anti-rat ALCAM/CD166 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1572-ST Lyophilized recombinant rat ALCAM/CD166 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ1572-CA Goat anti-rat ALCAM/CD166 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1572-DA Biotinylated goat anti-rat ALCAM/CD166 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1572-ST Lyophilized recombinant rat ALCAM/CD166 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ1572-CA Goat anti-rat ALCAM/CD166 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1572-CA Goat anti-rat ALCAM/CD166 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1572-DA Biotinylated goat anti-rat ALCAM/CD166 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) EZ1572-DA Biotinylated goat anti-rat ALCAM/CD166 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1572-ST Lyophilized recombinant rat ALCAM/CD166 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EZ1572-ST Lyophilized recombinant rat ALCAM/CD166 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Storage Conditions

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Calculated Molecular Weight

65022 MW

Specificity

Natural and recombinant rat ALCAM

Gene Name Synonym

CD166 antigen

Subcellular Location

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. Colocalizes with CD6 and the TCR/CD3 complex at the immunological synapse. .

Protein Name

CD166 antigen

Sample Data

Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.004 0.064 0.118 0.215 0.450 0.798 1.578 2.017 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.004 0.064 0.118 0.215 0.450 0.798 1.578 2.017 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.004 0.064 0.118 0.215 0.450 0.798 1.578 2.017 O.D. 0.004 0.064 0.118 0.215 0.450 0.798 1.578 2.017

Material Required But Not Provided

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)
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