Anti-Human Activated Leukocyte Cell Adhesion Molecule (ALCAM)
Purified No Carrier Protein
(CD166, MEMD, SC-1/DM-GRASP/BEN, KG-CAM)
Monoclonal Antibody
| Prod. No.: | A381 |
| Clone: | 105901 |
| Isotype: | Mouse IgG1 |
| Pkg. Size: | 1 mg, 500 µg |
| Storage: | -20°C to -70°C Detailed storage instructions below. |
Description
Specificity:
Mouse Anti-Human Activated Leukocyte Cell Adhesion Molecule (ALCAM) (Clone 105901) recognizes an epitope on Human ALCAM. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype. This antibody does not cross-react with rhBCAM, rmOCAM, rhMCAM and rhEpCAM.
Background:
This protein is a member of the immunoglobulin gene superfamily expressed by leukocytes after cell activation and thymic epithelial cells. In the nomenclature of CD antigens this protein has been given the designation CD166. ALCAM is a ligand for the lymphocyte antigen CD6 but also engages in homophilic interactions. Mouse ALCAM shows an overall identity of 93 % with the human protein. ALCAM has been identified independently as MEMD. mRNA transcripts encoding murine ALCAM are expressed in the brain, lung, liver, and the kidney, as well as by activated leukocytes and a number of cell lines. A rat monoclonal antibody specific for murine ALCAM binds to both activated CD4(+) and CD8(+) T-cells. The antigen is expressed on natural killer cells, thymocytes, platelets, B-cells and T-cells after cell activation, eosinophils, fibroblasts, endothelial cells, keratinocytes, monocytes after cell activation, neurons. The ligand for CD166 is CD6.
Host Species
Mouse
Immunogen
NS0-Derived Recombinant Human ALCAM Extracellular Domain
Formulation
This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.
Reconstitution
This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1.0 ml. This will result in a final antibody concentration of 500 µg/ml.
Endotoxin
<0.1 EU/µg as determined by the LAL method
Storage and Stability
The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.
Reported Applications
Indirect ELISA, WB, FA
Indirect ELISA: This antibody can be used in an indirect detection ELISA at 0.5 - 1.0 µg/ml with a suitable second step reagent such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (
Leinco Prod. No.: M114). The detection sensitivity of this indirect ELISA for Human ALCAM is approximately 2 ng/well.
Western Blotting: To detect Human ALCAM this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (
Leinco Prod. No.: M114) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (
Leinco Prod. No.: T343). The detection limit for Human ALCAM is 25 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used.
Functional Assay: This antibody shows inhibition in the HUT-78 adhesion assay. The adhesion of human HUT-78 cells (5 x 104 cells/well) to 10 μg/mL immobilized rhCD6/Fc (100 μL/well) was maximally inhibited (80 - 100%) following pre-treatment with antibody for 1 hour using 100 μL of a 5 μg/mL antibody solution per well.
Each investigator should determine their own optimal working dilution for specific applications.
Country of Origin
USA
Related Products
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
