Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Adhesion Blockade

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human ALCAM/CD166
Trp28-Ala526
Accession # AAB59499

Specificity

Detects human ALCAM/CD166 in direct ELISAs and Western blots. In direct ELISAs, approximately 25% cross-reactivity with recombinant mouse ALCAM/CD166 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human ALCAM/CD166 Antibody

Detection of Human ALCAM/CD166 by Western Blot

Detection of Human ALCAM/CD166 by Western Blot

ST6GAL1 targeting decreases levels of a subset of N-glycoproteins that are known BTIC regulators.(A) Schematic of proteomic analysis of D456 BTICs with and without ST6GAL1 KD (n = 4 for each group of shNT, sh32, and sh33). IB with samples independent of the proteomic analysis verified that successful targeting ST6GAL1 resulted in decreased (B) PDGFRB, (C) ALCAM, and (D) NRP1 protein. (E) Schematic of pulldown using SNA-bound Agarose beads. (F) SNA pulldown and protein A/G bound agarose beads as a control demonstrated that PDGFRB, ALCAM, and NRP1 were targets for alpha 2,6 sialylation. (G) SNA pulldown of D456 PDX cells with ST6GAL1 KD compared with NT, illustrating differential pulldown of PDGFRB. (H) PDGF-BB–induced (10 minutes) activation of PDGFRB in D456 GBM PDX cells with ST6GAL1 KD compared with NT; IB for p-PDGFRB and total PDGFRB. The experiments were repeated in at least 3 independent biological replicates. Data from 1 representative experiment are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36345944), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ALCAM/CD166 by Western Blot

Detection of Human ALCAM/CD166 by Western Blot

ST6GAL1 targeting decreases levels of a subset of N-glycoproteins that are known BTIC regulators.(A) Schematic of proteomic analysis of D456 BTICs with and without ST6GAL1 KD (n = 4 for each group of shNT, sh32, and sh33). IB with samples independent of the proteomic analysis verified that successful targeting ST6GAL1 resulted in decreased (B) PDGFRB, (C) ALCAM, and (D) NRP1 protein. (E) Schematic of pulldown using SNA-bound Agarose beads. (F) SNA pulldown and protein A/G bound agarose beads as a control demonstrated that PDGFRB, ALCAM, and NRP1 were targets for alpha 2,6 sialylation. (G) SNA pulldown of D456 PDX cells with ST6GAL1 KD compared with NT, illustrating differential pulldown of PDGFRB. (H) PDGF-BB–induced (10 minutes) activation of PDGFRB in D456 GBM PDX cells with ST6GAL1 KD compared with NT; IB for p-PDGFRB and total PDGFRB. The experiments were repeated in at least 3 independent biological replicates. Data from 1 representative experiment are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36345944), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ALCAM/CD166 by Western Blot

Detection of Human ALCAM/CD166 by Western Blot

ST6GAL1 targeting decreases levels of a subset of N-glycoproteins that are known BTIC regulators.(A) Schematic of proteomic analysis of D456 BTICs with and without ST6GAL1 KD (n = 4 for each group of shNT, sh32, and sh33). IB with samples independent of the proteomic analysis verified that successful targeting ST6GAL1 resulted in decreased (B) PDGFRB, (C) ALCAM, and (D) NRP1 protein. (E) Schematic of pulldown using SNA-bound Agarose beads. (F) SNA pulldown and protein A/G bound agarose beads as a control demonstrated that PDGFRB, ALCAM, and NRP1 were targets for alpha 2,6 sialylation. (G) SNA pulldown of D456 PDX cells with ST6GAL1 KD compared with NT, illustrating differential pulldown of PDGFRB. (H) PDGF-BB–induced (10 minutes) activation of PDGFRB in D456 GBM PDX cells with ST6GAL1 KD compared with NT; IB for p-PDGFRB and total PDGFRB. The experiments were repeated in at least 3 independent biological replicates. Data from 1 representative experiment are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36345944), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ALCAM/CD166 by Western Blot

Detection of Human ALCAM/CD166 by Western Blot

ST6GAL1 targeting decreases levels of a subset of N-glycoproteins that are known BTIC regulators.(A) Schematic of proteomic analysis of D456 BTICs with and without ST6GAL1 KD (n = 4 for each group of shNT, sh32, and sh33). IB with samples independent of the proteomic analysis verified that successful targeting ST6GAL1 resulted in decreased (B) PDGFRB, (C) ALCAM, and (D) NRP1 protein. (E) Schematic of pulldown using SNA-bound Agarose beads. (F) SNA pulldown and protein A/G bound agarose beads as a control demonstrated that PDGFRB, ALCAM, and NRP1 were targets for alpha 2,6 sialylation. (G) SNA pulldown of D456 PDX cells with ST6GAL1 KD compared with NT, illustrating differential pulldown of PDGFRB. (H) PDGF-BB–induced (10 minutes) activation of PDGFRB in D456 GBM PDX cells with ST6GAL1 KD compared with NT; IB for p-PDGFRB and total PDGFRB. The experiments were repeated in at least 3 independent biological replicates. Data from 1 representative experiment are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36345944), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human ALCAM/CD166 Antibody

Application
Recommended Usage

Adhesion Blockade

The adhesion of HuT 78 human cutaneous T cell lymphoma cells (5 x 104 cells/well) to immobilized Recombinant Human CD6 Fc Chimera (Catalog # 627-CD, 10 µg/mL, 100 µL/well) was maximally inhibited (80-100%) by 25 µg/mL of the antibody.

Western Blot

0.1 µg/mL
Sample: Recombinant Human ALCAM/CD166 Fc Chimera (Catalog # 656-AL)

Reviewed Applications

Read 1 review rated 5 using AF656 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

  1. Bowen, M.A. et al. (1995) J. Exp. Med. 181:2213.
  2. Aruffo, A. et al. (1997) Immunol. Today 18:498.
  3. 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

Additional ALCAM/CD166 Products

Product Documents for Human ALCAM/CD166 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human ALCAM/CD166 Antibody

For research use only

Citations for Human ALCAM/CD166 Antibody

Customer Reviews for Human ALCAM/CD166 Antibody (1)

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Customer Images


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  • Human ALCAM/CD166 Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Embryonic spinal cord
    Species: Mouse
    Verified Customer | Posted 05/08/2019
    E12.5 CD1 Mouse Spinal cord. In blue, the floor plate is labelled using Alcam/CD166 antibody.
    Human ALCAM/CD166 Antibody AF656

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