HA722755

NCAM1 / CD56 Recombinant Rabbit Monoclonal Antibody [PSH06-82]

cat.: HA722755

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Cynomolgus monkey, Pig
Applications:	WB, IHC-P, IHC-Fr, IF-Cell, FC, IF-Tissue
Clonality:	Monoclonal
Clone number:	PSH06-82

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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, Neuro-2a cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, mouse brain tissue, rat brain tissue, SH-SY5Y, Neuro-2a, C6, human peripheral blood lymphocytes.
Subcellular location:	Cell membrane.
Recommended Dilutions: WB IHC-P IHC-Fr IF-Cell FC IF-Tissue	1:3,000 1:1,000 1:500 1:100-1:1,000 1:1,000 1:100
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

	Fig1: Application: IHC-Fr Species: Mouse Site: Cerebral cortex Sample: Frozen section Antibody concentration: 1:500 Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.
	Fig2: Application: IHC-Fr Species: Rat Site: Cerebral cortex Sample: Frozen section Antibody concentration: 1:500 Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.
	Fig3: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-NCAM1 / CD56 antibody (HA722755) 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 (HA722755) 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.

	Fig4: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-NCAM1 / CD56 antibody (HA722755) 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 (HA722755) 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.
	Fig5: Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling NCAM1 / CD56 with Rabbit anti-NCAM1 / CD56 antibody (HA722755) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722755, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig6: Western blot analysis of NCAM1 / CD56 on different lysates with Rabbit anti-NCAM1 / CD56 antibody (HA722755) 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: Neuro-2a cell lysate (20 µg/Lane) Lane 4: C6 cell lysate (20 µg/Lane) Lane 5: Mouse brain tissue lysate (20 µg/Lane) Lane 6: Rat brain tissue lysate (20 µg/Lane) Predicted band size: 95 kDa Observed band size: 120-250 kDa Exposure time: Lane 1-3: 3 minutes; Lane 4-6: 30 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 (HA722755) 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.

Fig7: Immunocytochemistry analysis of SH-SY5Y cells labeling NCAM1 / CD56 with Rabbit anti-NCAM1 / CD56 antibody (HA722755) 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 (HA722755) 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.

Fig8: Immunocytochemistry analysis of Neuro-2a cells labeling NCAM1 / CD56 with Rabbit anti-NCAM1 / CD56 antibody (HA722755) 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 (HA722755) 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.

	Fig9: Immunocytochemistry analysis of C6 cells labeling NCAM1 / CD56 with Rabbit anti-NCAM1 / CD56 antibody (HA722755) 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 (HA722755) 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.
	Fig10: 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 (HA722755, 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).
	Fig11: 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 (HA722755, 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).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.