NCAM1 / CD56 Recombinant Rabbit Monoclonal Antibody [PSH06-53]
cat.: HA722756
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH06-53 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 95 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human NCAM1 aa 1-739. |
| Positive control: | SH-SY5Y cell lysate, C6 cell lysate, Rat brain tissue lysate, human brain tissue, rat brain tissue, SH-SY5Y, C6, human peripheral blood lymphocytes. |
| Subcellular location: | Cell membrane. |
| Recommended Dilutions:
WB IHC-P IF-Cell FC IP |
1:3,000 1:1,000 1:100-1:1,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P13591 Human | P13595 Mouse | P13596 Rat |
| Alternative names: | antigen MSK39 identified by monoclonal 5.1H11 antigen recognized by monoclonal 5.1H11 CD56 cell adhesion molecule, neural, 1 MSK 39 MSK39 N-CAM-1 NCAM 1 NCAM NCAM C NCAM-1 NCAM1 NCAM1_HUMAN NCAMC Neural cell adhesion molecule 1 Neural cell adhesion molecule NCAM OTTHUMP00000235666 |
Images
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Fig1:
Western blot analysis of NCAM1 / CD56 on different lysates with Rabbit anti-NCAM1 / CD56 antibody (HA722756) at 1/3,000 dilution. Lane 1: SH-SY5Y cell lysate (20 µg/Lane) Lane 2: HeLa cell lysate (negative) (20 µg/Lane) Lane 3: C6 cell lysate (20 µg/Lane) Lane 4: Rat brain tissue lysate (20 µg/Lane) Predicted band size: 95 kDa Observed band size: 120-250 kDa Exposure time: 5 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 (HA722756) at 1/3,000 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:
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-NCAM1 / CD56 antibody (HA722756) at 1/1,000 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 (HA722756) at 1/1,000 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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Fig3:
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-NCAM1 / CD56 antibody (HA722756) at 1/1,000 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 (HA722756) at 1/1,000 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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Fig4:
Immunocytochemistry analysis of SH-SY5Y cells labeling NCAM1 / CD56 with Rabbit anti-NCAM1 / CD56 antibody (HA722756) at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-NCAM1 / CD56 antibody (HA722756) at 1/1,000 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 (M1305-2, 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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Fig5:
Immunocytochemistry analysis of C6 cells labeling NCAM1 / CD56 with Rabbit anti-NCAM1 / CD56 antibody (HA722756) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-NCAM1 / CD56 antibody (HA722756) 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 (M1305-2, 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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Fig6:
Flow cytometric analysis of HeLa (left, negative) and SH-SY5Y (right, positive) cells labeling NCAM1 / CD56. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA722756, 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™ 594 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1122) 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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Fig7:
Flow cytometric analysis of human peripheral blood lymphocytes labeling NCAM1 / CD56. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA722756, 1μg/mL) (red). 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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Fig8:
NCAM1 / CD56 was immunoprecipitated from 0.2 mg SH-SY5Y cell lysate with HA722756 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722756 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SH-SY5Y cell lysate (input) Lane 2: HA722756 IP in SH-SY5Y cell lysate Lane 3: Rabbit IgG instead of HA722756 in SH-SY5Y cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 2 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.