Anti-NCAM1 antibody [EPR26939-108]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Perforin with ab324491 at 1/100 (10.21 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in subsets of human PBMCs (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab313779 Anti-NCAM1(CD56) rabbit monoclonal antibody was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution, followed by ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 (2 ug/ml) (Magenta).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Immunohistochemical analysis of paraffin-embedded Human appendix tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on stroma of human appendix (PMID : 27015206).The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum (PMID : 19246644).The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Immunohistochemical analysis of paraffin-embedded Human ovarian carcin tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human ovarian carcinoma.The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human astrocytoma.The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Human PBMC (human primary peripheral blood mononuclear cell) cells labelling NCAM1 with ab313779 at 1/500 (1.016 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing membranous staining in subsets of human PBMCs, which is not co-localized with CD3.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. Anti-human CD3 mouse monoclonal antibody (Alexa Fluor® 647) was used to counterstain tubulin at 1/200 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) cells labelling NCAM1 with ab313779 at 1/500 (1.016 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing membranous staining in No-GFP-CD16.NK-92 cell line.Negative control : MCF7 (PMID : 30050775).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt

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Flow Cytometry - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling NCAM1 with ab313779 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Cells are co-stained with CD3 conjugated to Alexa Fluor®647. Gated on viable cells.

• Flow Cyt

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Flow Cytometry - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Flow cytometric analysis of MCF7 (human breast adenocarcinoma epithelial cell, Left) - / No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) (Right) cells labelling NCAM1 with ab313779 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control : MCF7 (PMID : 30050775). Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling NCAM1 with ab313779 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human liver.The section was incubated with ab313779 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The 120, 140 and 180kDa bands are different isoforms as reported in the literature (PMID : 26288071).

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-NCAM1 antibody [EPR26939-108] (ab313779) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 120 kDa,140 kDa,180 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-NCAM1 antibody [EPR26939-108] (AB313779)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : MCF7 (PMID : 18772882).

The 120, 140 and 180kDa bands are different isoforms as reported in the literature (PMID : 26288071)

The identity of the lower MW bands at approximately 75 kDa and 50kDa are unknown.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

Exposure time : Lane 1 : 147 seconds; Lane 2, 3 : 92 seconds.

All lanes:

Western blot - Anti-NCAM1 antibody [EPR26939-108] (ab313779) at 1/1000 dilution

Lane 1:

No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92&reg;) cell with no GFP tag) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 120 kDa,140 kDa,180 kDa

false