Western blot shows lysates of A549 human lung carcinoma cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human ALCAM/CD166 Monoclonal Antibody (Catalog # MAB6563) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for ALCAM/CD166 at approximately 105 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of ALCAM in HEK293 Human Cell Line Transfected with Human ALCAM and eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with (A) human ALCAM or (B) irrelevant protein and eGFP was stained with Rabbit Anti-Human ALCAM Monoclonal Antibody (Catalog # MAB6563) followed by APC-conjugated Anti-Rabbit IgG Secondary Antibody (F0111). Quadrant markers were set based on control antibody staining (MAB1050). Staining was performed using our Staining Membrane-associated Proteins protocol.

ALCAM/CD166 in A431 Human Cell Line.

ALCAM/CD166 was detected in immersion fixed A431 human epithelial carcinoma cell line using Rabbit Anti-Human ALCAM/CD166 Monoclonal Antibody (Catalog # MAB6563) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cell surface.Staining was performed using our Fluorescent ICC Staining of Cells on Coverslips Protocol.

Applications for Human ALCAM/CD166 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 Human Cell Line Transfected with Human ALCAM and eGFP

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed A431 human epithelial carcinoma cell line

Western Blot

2 µg/mL
Sample: A549 human lung carcinoma cell line and U2OS human osteosarcoma cell line

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: ALCAM/CD166

ALCAM, activated leukocyte cell adhesion molecule, is a type I membrane glycoprotein and a member of the immunoglobulin supergene family. It is also known as CD166, MEMD, SC-1/DM-GRASP/BEN in the chicken, and KG-CAM in the rat. ALCAM is expressed on thymic epithelial cells, activated B and T cells, and monocytes. ALCAM can bind itself homotypically and is also capable of binding CD6, NgCAM, and other, as of yet, unidentified brain proteins. The ALCAM/CD6 interaction may be involved in T cell development and T cell regulation. Additionally, ALCAM/CD6 and ALCAM/NgCAM interactions may play roles in the nervous system. ALCAM has also been observed to be upregulated on highly metastasizing melanoma cell lines and may play a role in tumor migration. ALCAM is a 583 amino acid (aa) protein consisting of a 27 aa signal peptide, a 500 aa extracellular domain, a 24 aa transmembrane domain and a 32 aa cytoplasmic domain. The extracellular domain of ALCAM contains 5 Ig-like domains.

References

Bowen, M.A. et al. (1995) J. Exp. Med. 181:2213.
Aruffo, A. et al. (1997) Immunol. Today 18:498.
Degen, W.G. et al. (1998) Am. J. Pathol. 152:805.

Long Name

Activated Leukocyte Cell Adhesion Molecule

Alternate Names

CD166

Entrez Gene IDs

214 (Human); 11658 (Mouse); 101867493 (Cynomolgus Monkey)

Gene Symbol

ALCAM

UniProt

Q13740

Additional ALCAM/CD166 Products

Product Documents for Human ALCAM/CD166 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human ALCAM/CD166 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human ALCAM/CD166 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB6563

Key Product Details

Species Reactivity

Validated:

Applications

Label

Antibody Source

View all ALCAM/CD166 Antibodies »

Product Summary for Human ALCAM/CD166 Antibody

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human ALCAM/CD166 Antibody

Detection of Human ALCAM/CD166 by Western Blot.