CD166 Mouse Monoclonal Antibody [5-5]
cat.: EM1902-36
| Product Type: | Mouse monoclonal IgG1, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IHC-P, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | 5-5 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 1%BSA, 50%Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 2ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 65 kDa. |
| Isotype: | IgG1 |
| Immunogen: | Recombinant protein within human CD166 aa 1-300. |
| Positive control: | PC-3M cell lysate, A549 cell lysate, SHSY5Y cell lysate, A431cells, Huvec celles, human liver tissue, THP-1 cells. |
| Subcellular location: | Cell membrane, Cell projection, Membrane, Secreted |
| Recommended Dilutions:
WB IHC-P IF-Cell FC |
1:500-1:1,000 1:50-1:200 1:50-1:100 1:50-1:100 |
| Uniprot #: | SwissProt: Q13740 Human |
| Alternative names: | Activated leukocyte cell adhesion molecule ALCAM ALCAM protein CD 166 CD166 CD166 antigen CD166_HUMAN FLJ3851 FLJ38514 MEMD MGC71733 |
Images
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Fig1:
Western blot analysis of CD166 on different lysates with Mouse anti-CD166 antibody (EM1902-36) at 1/1,000 dilution. Lane 1: A549-WT cell lysate Lane 2: A549-KD CD166 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 65 kDa Observed band size: 100 kDa Exposure time: 2 minutes 10 seconds; ECL: K1802; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1902-36) at 1/1,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
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 |
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Fig3: 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-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue). |
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Fig4: 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). |
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Fig5: 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. |
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Fig6: 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; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.