CD146 Recombinant Rabbit Monoclonal Antibody [JB42-35]
cat.: ET7107-91
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P, FC |
| Clonality: | Monoclonal |
| Clone number: | JB42-35 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 72 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within C-terminal human CD146 (Cytoplasmic). |
| Positive control: | A375 cell lysate, HeLa cell lysate, HUVEC cell lysate, HepG2 cell lysate, Mouse lung tissue lysate, A375, human tonsil tissue. |
| Subcellular location: | Membrane. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC |
1:1,000 1:100 1:200 1:1,000 |
| Uniprot #: | SwissProt: P43121 Human | Q8R2Y2 Mouse | Q9EPF2 Rat |
| Alternative names: | A32 antigen CD 146 CD146 CD146 antigen Cell surface glycoprotein MUC18 Cell surface glycoprotein P1H12 Gicerin Mcam Melanoma adhesion molecule Melanoma associated antigen A32 Melanoma associated antigen MUC18 Melanoma associated glycoprotein MUC18 Melanoma cell adhesion molecule Melanoma-associated antigen A32 Melanoma-associated antigen MUC18 MelCAM MUC 18 MUC18 MUC18_HUMAN S endo 1 S endo 1 endothelial associated antigen S-endo 1 endothelial-associated antigen |
Images
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Fig1:
Western blot analysis of CD146 on different lysates with Rabbit anti-CD146 antibody (ET7107-91) at 1/1,000 dilution. Lane 1: A375 cell lysate (20 µg/Lane) Lane 2: HeLa cell lysate (20 µg/Lane) Lane 3: HUVEC cell lysate (20 µg/Lane) Lane 4: HepG2 cell lysate (20 µg/Lane) Lane 5: Mouse lung tissue lysate (40 µg/Lane) Predicted band size: 72 kDa Observed band size: 120 kDa Exposure time: 14 seconds; ECL: K1801; 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 (ET7107-91) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of CD146 on different lysates with Rabbit anti-CD146 antibody (ET7107-91) at 1/1,000 dilution. Lane 1: HAP1-parental cell lysate Lane 2: HAP1-CD146 KD cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 72 kDa Observed band size: 120 kDa Exposure time: 180 seconds; ECL: K1801; 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 (ET7107-91) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunocytochemistry analysis of A375 cells labeling CD146 with Rabbit anti-CD146 antibody (ET7107-91) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD146 antibody (ET7107-91) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-CD146 antibody (ET7107-91) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-91) at 1/200 dilution for 1 hour 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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Fig5:
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-CD146 antibody (ET7107-91) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-91) at 1/200 dilution for 1 hour 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 A375 cells labeling CD146. Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-91, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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