PRODUCT CODE: EM1902-36

CD166 Mouse Monoclonal Antibody [5-5] (EM1902-36)

Applications

  • WB

  • IHC-P

  • ICC

  • FC

REACTIVITY

  • Human

Western blot analysis of CD166 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1902-36, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: PC-3M cell lysate<br />
Lane 2: A549 cell lysate<br />
Lane 3: SHSY5Y cell lysate
  • Western blot analysis of CD166 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1902-36, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: PC-3M cell lysate<br />
Lane 2: A549 cell lysate<br />
Lane 3: SHSY5Y cell lysate
  • ICC staining of CD166 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1902-36, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of CD166 in Huvec cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1902-36, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-CD166 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-36, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of CD166 was done on THP-1 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1902-36, 1/100) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-Mouse IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; blcak).
Western blot analysis of CD166 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1902-36, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: PC-3M cell lysate
Lane 2: A549 cell lysate
Lane 3: SHSY5Y cell lysate

Applications

  • WB

  • IHC-P

  • ICC

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

CD166 Mouse Monoclonal Antibody [5-5] (EM1902-36)

Immunogen

Recombinant protein within human cd166 aa 1-300.

Host

Mouse

Positive Control

PC-3M cell lysate, A549 cell lysate, SHSY5Y cell lysate, A431cells, Huvec celles, human liver tissue, THP-1 cells.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

5-5

PROPERTIES

Form

Liquid

Storage Condition

Store at +4Á¾ after thawing. Aliquot store at -20Á¾. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 1%BSA, 50%Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

Predicted band size: 65 kDa.

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:500-1:1,000

  • IHC-P

  • 1:50-1:200

  • ICC

  • 1:50-1:100

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

CD166

SYNONYMS

Activated leukocyte cell adhesion molecule antibody; ALCAM antibody; ALCAM protein antibody; CD 166 antibody; CD166 antibody; CD166 antigen antibody; CD166_HUMAN antibody; FLJ3851 antibody; FLJ38514 antibody; MEMD antibody; MGC71733 antibody

SUBCELLULAR LOCATION

Cell membrane, Cell projection, Membrane, Secreted

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 . Contributes to the formation and maturation of the immunological synapse via its interactions with CD6 . Mediates homotypic interactions with cells that express ALCAM . Required for normal hematopoietic stem cell engraftment in the bone marrow . 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. Mediates outgrowth and pathfinding for retinal ganglion cell axons (By similarity).